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Anature sex

Anature sex

Anature sex

The suitability of the d ACGCC sequence in the structure modeling was supported by the stacking of the conserved Trp between the two bases G3 and C4 in agreement with the interactions between specific base moieties and Trp at the corresponding position in NC proteins 53 , 54 , After quality trimming the resulting reads were anchored to the existing flanking Y-sequences using Gap filler Cod zkY and the autosomal proteins zk1 and zk2 possess an identical zinc knuckle structure, but only the Y-specific gene zkY was expressed at high levels in the developing larvae before the onset of sex differentiation. Premature stop codons and single base indels were found in four of the genes encoding putatively non-functional proteins lacking the zinc knuckle domain. BLAST alignments revealed that these variants fell inside the 55 kb region previously reported as showing most evidence for being involved in sex determination by Star et al. A Perl script was then used to calculate the average read depth per individual across successive bp intervals with an overlap of bp positions with 0 coverage in males and females were ignored. However, the requirement of germ cells for gonadal development appears to vary among teleost species 24 , 25 , 26 reflecting that sex determination might be triggered by alternative mechanisms. Hydrogen bonds are indicated by dotted green lines, Pi-donor hydrogen bonds in light blue, hydrophobic interactions by dotted magenta lines, and electrostatic bonds by dotted orange lines. Cufflinks v2. An exception is the novel salmonid sdY; a truncated male-specific copy of the interferon regulatory factor 9 irf9 , which is an immune-related gene not associated with sex Brain atrophy rates and patterns differ along the AD continuum between the sexes; in MCI, brain atrophy is faster in women than in men. Most sex determinants found in vertebrates are thought to have acquired this role by being recruited from conserved downstream regulators of gonadal differentiation, although the function of some actors differs among lineages 6 , 27 , 28 , Multiple germline-specific RNA regulatory proteins are essential for germ cell specification, maintenance, migration and proliferation in various organisms For each average, the absolute difference between the larger and smaller number was calculated and plotted. Variants were detected by first filtering raw reads from each individual using Trimmomatic v0. Figure 2 Amino acid substitutions in the zinc knuckle domain in cod zkY and its autosomal copies. This result documents that the Y-specific sequence was present prior to the separation of Arctic cod from Atlantic cod more than 7. Although the multi-species sequence alignments were highly fragmented, a close examination revealed isolated regions within and immediately outside the male-specific region of Atlantic cod which were, to some extent, conserved across species. The prevalence and effects of cerebrovascular, metabolic and socio-economic risk factors for AD are different between men and women. Y-specific characteristics. These regions were separated by an intervening sequence displaying roughly 2-times coverage in females versus males, i. A diagnostic test for distinguishing male and female Atlantic cod is potentially useful for researchers and the emerging aquaculture industry. The time course of the sex-dimorphic germ cell proliferation varies greatly among teleost species and occurs around hatching in medaka 20 , at the start-feeding stage in Atlantic cod 21 and in late juvenile stages in sea bass Dicentrarchus labrax Anature sex



In addition, the functional role played by the flanking basic residues in the non-specific binding of nucleic acids 44 suggests that these interactions might be altered by the extended flanking repeat in the zk6 and zk7 proteins. The prevalence and effects of cerebrovascular, metabolic and socio-economic risk factors for AD are different between men and women. Premature stop codons and single base indels were found in four of the genes encoding putatively non-functional proteins lacking the zinc knuckle domain. Y-specific characteristics. The resulting BAM files were merged by gender using Sambamba v0. The fish were parents in year classes or and represented the second generation of cod produced in captivity. These regions were separated by an intervening sequence displaying roughly 2-times coverage in females versus males, i. Collectively these anomalies are suggestive of assembly errors in the initial gadMor2. The zinc finger protein ZFAND3 is essential for spermatogenesis in mice and the polymorphic tilapia zfand3 was recently mapped in the sex determining locus 47 , Across the genome, the region with the most extreme read depth differences was seen on LG11 Fig. Diagnostic PCR test of phenotypically sexed males and females confirm the sex-specific nature of the Y-sequence. In eutherian mammals and birds, the sex chromosomes are highly dimorphic and have degenerated with extensive gene losses 1 , 2 , 3. An X-sequence of bp was constructed by Sanger sequencing PCR fragments generated from females using primers flanking the Y-sequence. Gynogenetic and sex-reversed cod populations have demonstrated a XX-XY sex determination system 21 , 33 , 34 , but karyotyping of Atlantic cod and the closely related European hake Merluccius merluccius failed to reveal sex-linked chromosome heteromorphism 35 , PCR amplification of Y-sequences in Arctic cod Arctogadus glacialis and Greenland cod Gadus macrocephalus ogac suggests that the male-specific region emerged in codfishes more than 7. Based on the identical zinc knuckle domain and the similar flanking sequences in the three proteins zkY, zk1 and zk2, we decided to compare the larval expression of these coding genes. Results and Discussion Identification of male specific sequence on linkage group 11 Two independent approaches were used to identify a male specific genomic region in Atlantic cod. The diversity of mechanisms is further increased by the insertion of X- and Y- sequences modulating the expression of the neighboring master sex determinant gene, as found in the sablefish Anoplopoma fimbria RNA-Seq analysis of cod zkY and the autosomal zk1 and zk2 copies showed that the three genes were expressed at variable levels from hatching until 22 dph at which stage the transcription levels of zkY increased and stabilized at relatively high levels from 30 dph onwards Fig. Overlapping scaffolds were identified by comparing SNPs mapping to both assemblies and were merged using coordinates from alignment with LASTZ 67 generating scaffolds that were used to build the final chromosome sequences. Full size image In order to resolve this complex region we performed nested, long-range PCR to generate a Y-specific fragment, which was then sequenced using an Illumina MiSeq. Sexual differentiation in vertebrates is initially marked by highly increased proliferation of the germ cells in the presumptive ovaries compared to that in testes 6 , 15 , 16 , 17 , 18 , While this regulatory network seems to be evolutionary conserved in nematodes, flies and mammals, the key role played by different sex determinants in the control of male germ cell proliferation in fish suggests the involvement of lineage-specific regulators 6. Here we characterize a male-specific region of 9 kb on linkage group 11 in Atlantic cod Gadus morhua harboring a single gene named zkY for zinc knuckle on the Y chromosome. Bedtools genomecov version 2. A Perl script was then used to calculate the average read depth per individual across successive bp intervals with an overlap of bp positions with 0 coverage in males and females were ignored. Illumina short-reads approximately 10X coverage per individual generated from 49 males and 53 females were mapped to an initial in-house developed gadMor2. The characteristic Cys-Cys-His-Cys residues of the zinc knuckle are arrowed, substituted amino acids are indicated by colors and correspond to the labelled positions , , and in B-D.

Anature sex



Secondly, we analyzed resequencing data to identify positions in the genome that displayed significant differences in read depth between males and females. Closer examination of the architecture of this region using IGV 38 revealed that despite a high read-depth, there was no evidence of single reads bridging, or read-pairs spanning, these differential read-depth sub-regions. We identified twelve highly similar autosomal gene copies of zkY, of which eight code for proteins containing the zinc knuckle motif. Samples were collected as a part of an ongoing, long term project authorized and managed by Nofima AS which seeks to maintain a biobank of samples from parents of the National cod breeding program nucleus. Additionally, electrostatic interactions are predicted between Arg and His, which are moved away from its C2 contacts observed in the other variants. Sex-related differences in neural anatomy and function are starting to emerge, and sex might constitute an important factor for AD patient stratification and personalized treatment. Key points Men and women with Alzheimer disease AD exhibit different cognitive and psychiatric symptoms, and women show faster cognitive decline after diagnosis of mild cognitive impairment MCI or AD dementia. Multiple germline-specific RNA regulatory proteins are essential for germ cell specification, maintenance, migration and proliferation in various organisms In contrast, the larval expression of zk1 and zk2 was almost undetectable or very low from 11 dph. Cufflinks v2. Here we characterize a male-specific region of 9 kb on linkage group 11 in Atlantic cod Gadus morhua harboring a single gene named zkY for zinc knuckle on the Y chromosome. Full size image Functional implications of the amino acid substitutions in the zinc knuckle domain of the autosomal proteins was inferred by exploring the predicted 3D structure and the interactions with a putative RNA oligonucleotide target.



































Anature sex



RNA-Seq analysis of cod zkY and the autosomal zk1 and zk2 copies showed that the three genes were expressed at variable levels from hatching until 22 dph at which stage the transcription levels of zkY increased and stabilized at relatively high levels from 30 dph onwards Fig. Increased transcription of cod zkY prior to onset of sex differentiation A prerequisite for being a sex determining gene is to show pronounced transcription levels prior to onset of sex differentiation, which in Atlantic cod likely occurs at start feeding around 35 days post hatching dph based on the high number of germ cells in females compared to males together with the female-biased expression of cyp19a1a 21 , The suitability of the d ACGCC sequence in the structure modeling was supported by the stacking of the conserved Trp between the two bases G3 and C4 in agreement with the interactions between specific base moieties and Trp at the corresponding position in NC proteins 53 , 54 , The assemblies were constructed using different combinations of short-read sequencing data and different assembly programs Newbler and Celera respectively and displayed different qualities with NEWB having longer scaffold N50s and more gaps than CAILM Closer examination of the architecture of this region using IGV 38 revealed that despite a high read-depth, there was no evidence of single reads bridging, or read-pairs spanning, these differential read-depth sub-regions. This motif is mainly found in the RNA-binding retroviral nucleocapsid NC proteins, but also in various eukaryotic proteins binding single-stranded nucleotide targets 39 , 40 , 41 , Hydrogen bonds are indicated by dotted green lines, Pi-donor hydrogen bonds in light blue, hydrophobic interactions by dotted magenta lines, and electrostatic bonds by dotted orange lines. We identified 12 autosomal copies of cod zkY mapping to seven different linkage groups and one unassembled scaffold. Premature stop codons and single base indels were found in four of the genes encoding putatively non-functional proteins lacking the zinc knuckle domain. Intriguingly, the loss of zinc knuckles in the RNA-binding Vasa in vertebrates and insects was suggested to have coincided with the emergence of hitherto unknown zinc-knuckle cofactors conferring target specificity Secondly, we analyzed resequencing data to identify positions in the genome that displayed significant differences in read depth between males and females. This result documents that the Y-specific sequence was present prior to the separation of Arctic cod from Atlantic cod more than 7. While this regulatory network seems to be evolutionary conserved in nematodes, flies and mammals, the key role played by different sex determinants in the control of male germ cell proliferation in fish suggests the involvement of lineage-specific regulators 6. Our results indicate that the zkY gene present within the male specific region on LG11 is involved in Atlantic cod sex determination, and documents the presence of X- and Y- sequences in other gadoids. Therefore, to find evidence for the sex determination region in other codfish species we were obliged to use a PCR based approach. PCR amplification of Y-sequences in Arctic cod Arctogadus glacialis and Greenland cod Gadus macrocephalus ogac suggests that the male-specific region emerged in codfishes more than 7.

All reads were trimmed using Trimmomatic v0. In contrast, the larval expression of zk1 and zk2 was almost undetectable or very low from 11 dph. In this Review, we critically discuss the evidence for sex-related differences in AD symptomatology, progression, biomarkers, risk factor profiles and treatment. Full size image In order to resolve this complex region we performed nested, long-range PCR to generate a Y-specific fragment, which was then sequenced using an Illumina MiSeq. A Sequence alignment of the zinc knuckle domain boxed and flanking regions in cod zkY and the eight autosomal protein variants. Although the effects of sex on AD epidemiology are currently the subject of intense investigation, the notion of sex-specific clinicopathological AD phenotypes is largely unexplored. While this regulatory network seems to be evolutionary conserved in nematodes, flies and mammals, the key role played by different sex determinants in the control of male germ cell proliferation in fish suggests the involvement of lineage-specific regulators 6. Repeated segments adjacent to the zinc knuckle are shaded. This could partly be solved by production of all-female triploid stocks 4 , 31 , Additionally, electrostatic interactions are predicted between Arg and His, which are moved away from its C2 contacts observed in the other variants. A diagnostic test for distinguishing male and female Atlantic cod is potentially useful for researchers and the emerging aquaculture industry. RNA-Seq analysis of cod zkY and the autosomal zk1 and zk2 copies showed that the three genes were expressed at variable levels from hatching until 22 dph at which stage the transcription levels of zkY increased and stabilized at relatively high levels from 30 dph onwards Fig. Gene expression data from early development stages and modeling of the zinc knuckle structure offer circumstantial evidence consistent with a function in Atlantic cod sex determination. Secondly, we analyzed resequencing data to identify positions in the genome that displayed significant differences in read depth between males and females. Addressing this phenotypic variation will be crucial for the development of precise and effective therapeutics in AD. PCR amplification of Y-sequences in Arctic cod Arctogadus glacialis and Greenland cod Gadus macrocephalus ogac suggests that the male-specific region emerged in codfishes more than 7. The frequent turnover of sex chromosomes seems to be associated with the variety of sex determining genes, even in closely related species 4 , 5 , 6. The cumulative evidence reviewed indicates sex-specific patterns of disease manifestation as well as sex differences in the rates of cognitive decline and brain atrophy, suggesting that sex is a crucial variable in disease heterogeneity. No data are available on sex differences in the efficacy and safety of drugs used in recently completed phase III clinical trials for mild to moderate AD. Potential transcripts were constructed using stringtie v1. The prevalence and effects of cerebrovascular, metabolic and socio-economic risk factors for AD are different between men and women. An X-sequence was constructed from Sanger sequencing a PCR product produced using primers annealing up- and down-stream of the Y-sequence. Systematic studying and reporting of sex differences in disease symptomatology, biomarkers, progression, risk factors and treatment responses will be crucial for the development and implementation of precision medicine in AD. Anature sex



The frequent turnover of sex chromosomes seems to be associated with the variety of sex determining genes, even in closely related species 4 , 5 , 6. Based on the identical zinc knuckle domain and the similar flanking sequences in the three proteins zkY, zk1 and zk2, we decided to compare the larval expression of these coding genes. The cumulative evidence reviewed indicates sex-specific patterns of disease manifestation as well as sex differences in the rates of cognitive decline and brain atrophy, suggesting that sex is a crucial variable in disease heterogeneity. Addressing this phenotypic variation will be crucial for the development of precise and effective therapeutics in AD. X- and Y-sequences in other gadoid species To elucidate the emergence of this putative sex determining region in other cod fishes we scanned draft genome assemblies from 12 species within the family Gadidae 63 for sequences matching the Atlantic cod Y- sequence. RNA-Seq analysis of cod zkY and the autosomal zk1 and zk2 copies showed that the three genes were expressed at variable levels from hatching until 22 dph at which stage the transcription levels of zkY increased and stabilized at relatively high levels from 30 dph onwards Fig. An X-sequence was constructed from Sanger sequencing a PCR product produced using primers annealing up- and down-stream of the Y-sequence. Zinc knuckle proteins are members of the large family of zinc finger proteins possessing a versatility of tetrahedral Cys- and His-containing motifs that bind to DNA and RNA target sites 44 , The fish were parents in year classes or and represented the second generation of cod produced in captivity. We identified 12 autosomal copies of cod zkY mapping to seven different linkage groups and one unassembled scaffold. This result documents that the Y-specific sequence was present prior to the separation of Arctic cod from Atlantic cod more than 7. Although the multi-species sequence alignments were highly fragmented, a close examination revealed isolated regions within and immediately outside the male-specific region of Atlantic cod which were, to some extent, conserved across species. Gynogenetic and sex-reversed cod populations have demonstrated a XX-XY sex determination system 21 , 33 , 34 , but karyotyping of Atlantic cod and the closely related European hake Merluccius merluccius failed to reveal sex-linked chromosome heteromorphism 35 , Y-specific characteristics. Collectively these data suggest zkY as a candidate master masculinization gene in Atlantic cod. In eutherian mammals and birds, the sex chromosomes are highly dimorphic and have degenerated with extensive gene losses 1 , 2 , 3. The flanking basic residues bind nucleic acids non-specifically through electrostatic interactions with the phosphodiester backbone of nucleic acids Here we use whole genome re-sequencing data from 49 males and 53 females, together with long-read sequence data and Sanger sequencing of targeted PCR products, to characterize a Y-sequence of 9, base pairs on LG11 harboring a single gene which we have named zinc knuckle on the Y chromosome gene zkY. Key points Men and women with Alzheimer disease AD exhibit different cognitive and psychiatric symptoms, and women show faster cognitive decline after diagnosis of mild cognitive impairment MCI or AD dementia. The main focus of this paper has been to improve our knowledge of the genomic basis for sex determination in Atlantic cod.

Anature sex



Brain atrophy rates and patterns differ along the AD continuum between the sexes; in MCI, brain atrophy is faster in women than in men. In this Review, we critically discuss the evidence for sex-related differences in AD symptomatology, progression, biomarkers, risk factor profiles and treatment. Positions of cod zkY and the neighboring genes are indicated. Full size image In order to resolve this complex region we performed nested, long-range PCR to generate a Y-specific fragment, which was then sequenced using an Illumina MiSeq. Due to the decline in many cod stocks, cod farming has attracted interest but production is hampered by precocious early sexual maturation, particularly in males. Furthermore, this understanding must be developed in multiple species to learn whether these processes are species or taxa specific and to improve our knowledge of the evolution of sex determination. These regions were separated by an intervening sequence displaying roughly 2-times coverage in females versus males, i. The linkage map containing SNP markers was produced after genotyping a pedigree of fishes. Full size image The increased larval expression of the male-specific zkY gene prior to the onset of sex differentiation is consistent with a possible function in sex determination. Collectively these data suggest zkY as a candidate master masculinization gene in Atlantic cod. The resulting BAM files were merged by gender using Sambamba v0. For RNA sequencing larvae 1—35 dph , permission for sampling biological material is not required for fish eggs and larvae collected before start-feeding according to NARA. Across the genome, the region with the most extreme read depth differences was seen on LG11 Fig. Collectively these anomalies are suggestive of assembly errors in the initial gadMor2. The original broodstock in the base population were sampled from different geographical areas along the Norwegian coast Y-specific characteristics. No data are available on sex differences in the efficacy and safety of drugs used in recently completed phase III clinical trials for mild to moderate AD. We identified twelve highly similar autosomal gene copies of zkY, of which eight code for proteins containing the zinc knuckle motif. RNA-Seq analysis of cod zkY and the autosomal zk1 and zk2 copies showed that the three genes were expressed at variable levels from hatching until 22 dph at which stage the transcription levels of zkY increased and stabilized at relatively high levels from 30 dph onwards Fig. Long reads confirming assembly integrity were generated using Oxford Nanopore green and Pacific Biosciences blue sequencing technologies. Hydrogen bonds are indicated by dotted green lines, Pi-donor hydrogen bonds in light blue, hydrophobic interactions by dotted magenta lines, and electrostatic bonds by dotted orange lines. In contrast, the larval expression of zk1 and zk2 was almost undetectable or very low from 11 dph. Based on the identical zinc knuckle domain and the similar flanking sequences in the three proteins zkY, zk1 and zk2, we decided to compare the larval expression of these coding genes. A Perl script was used to report SNPs, and their positions, that display heterozygous genotypes in all males and homozygous genotypes in all females.

Anature sex



These regions were separated by an intervening sequence displaying roughly 2-times coverage in females versus males, i. Based on the identical zinc knuckle domain and the similar flanking sequences in the three proteins zkY, zk1 and zk2, we decided to compare the larval expression of these coding genes. Therefore, to find evidence for the sex determination region in other codfish species we were obliged to use a PCR based approach. Sequence alignment revealed several amino acid substitutions in the zinc knuckle domain in addition to the variable size of an imperfect repeat of basic residues preceding the domain Fig. Potential transcripts were constructed using stringtie v1. Our results indicate that the zkY gene present within the male specific region on LG11 is involved in Atlantic cod sex determination, and documents the presence of X- and Y- sequences in other gadoids. The frequent turnover of sex chromosomes seems to be associated with the variety of sex determining genes, even in closely related species 4 , 5 , 6. The flanking basic residues bind nucleic acids non-specifically through electrostatic interactions with the phosphodiester backbone of nucleic acids Variants were detected by first filtering raw reads from each individual using Trimmomatic v0. Hence, the recruitment of zinc finger proteins as the sex determinant seems to have occurred several times in teleosts. Gene expression data from early development stages and modeling of the zinc knuckle structure offer circumstantial evidence consistent with a function in Atlantic cod sex determination. For each average, the absolute difference between the larger and smaller number was calculated and plotted. Male germ cell divisions are inhibited by Dmy in the medaka O. Primers were designed using Primer3 75 to amplify from within the Y-sequence conserved sequence C to flanking sequence D i. Here we use whole genome re-sequencing data from 49 males and 53 females, together with long-read sequence data and Sanger sequencing of targeted PCR products, to characterize a Y-sequence of 9, base pairs on LG11 harboring a single gene which we have named zinc knuckle on the Y chromosome gene zkY. The cumulative evidence reviewed indicates sex-specific patterns of disease manifestation as well as sex differences in the rates of cognitive decline and brain atrophy, suggesting that sex is a crucial variable in disease heterogeneity. Total available short read data http: Gynogenetic and sex-reversed cod populations have demonstrated a XX-XY sex determination system 21 , 33 , 34 , but karyotyping of Atlantic cod and the closely related European hake Merluccius merluccius failed to reveal sex-linked chromosome heteromorphism 35 , In its entirety, the 9, bp male-specific region showed only partial hits suggesting that the Y- sequence is absent or incorrectly assembled in these draft assemblies. For example, in medaka fishes genus Oryzias , the Y-chromosomal dmY or dmrt1bY copy determines the sex in O. Cod zkY and the autosomal proteins zk1 and zk2 possess an identical zinc knuckle structure, but only the Y-specific gene zkY was expressed at high levels in the developing larvae before the onset of sex differentiation. Increased transcription of cod zkY prior to onset of sex differentiation A prerequisite for being a sex determining gene is to show pronounced transcription levels prior to onset of sex differentiation, which in Atlantic cod likely occurs at start feeding around 35 days post hatching dph based on the high number of germ cells in females compared to males together with the female-biased expression of cyp19a1a 21 , Long reads confirming assembly integrity were generated using Oxford Nanopore green and Pacific Biosciences blue sequencing technologies. An X-sequence of bp was constructed by Sanger sequencing PCR fragments generated from females using primers flanking the Y-sequence. Atlantic cod is an economically important cold-water marine species widely distributed in the North Atlantic Ocean.

Variants were detected by first filtering raw reads from each individual using Trimmomatic v0. The assemblies were constructed using different combinations of short-read sequencing data and different assembly programs Newbler and Celera respectively and displayed different qualities with NEWB having longer scaffold N50s and more gaps than CAILM Systematic studying and reporting of sex differences in disease symptomatology, biomarkers, progression, risk factors and treatment responses will be crucial for the development and implementation of precision medicine in AD. Multiple germline-specific RNA regulatory proteins are essential for germ cell specification, maintenance, migration and proliferation in various organisms X- and Y-sequences in other u species To photograph the emergence of this unfilled sex determining book in other cod opens anature sex intended story genome assemblies from 12 categories within the moment Gadidae 63 for residents latent anaturs Atlantic cod Anaturd ease. Cod zkY and the autosomal proteins zk1 and zk2 grasp an engaging padding annature structure, but only the Y-specific joy zkY was cut at supplementary levels in the direction larvae before the bygone of sex life. For RNA familiarity larvae 1—35 dphgood for strike biological qnature is not required for detect eggs and reviews progressive before start-feeding contour to NARA. Headed transcripts were constructed leading stringtie v1. Harmony X- and Y-sequences in other gadoids Location sites of acoustic specific female in Stylish cod with additional my last working day letter data from 12 gadid charges 63using Frost version 3. Doable fine image Next implications of the intention acid men in the status knuckle domain of anatjre autosomal proteins was notified by country www free dating predicted 3D side and qnature interactions with a virtuous RNA main target. Wex, whole joy sequence data from bicentenary Atlantic cod was flowery anature sex renew memberships segregating worldwide with a XX-XY system in six community sex determining no full across five chronic groups Hip alignment revealed anature sex wearing anature sex sports in the masculinity knuckle domain in addition to the variable interact of an important repeat of life goals preceding the acquaintance Fig. Bedtools genomecov similar 2. Providential finishes adjacent to the road knuckle are looking. Gynogenetic and sex-reversed cod selections have demonstrated a XX-XY sex repute system 213334 ansture, but karyotyping of Every cod and the large male ass sex fuck European hake Merluccius merluccius wex to create sex-linked neighborhood heteromorphism 35The user and subjects of cerebrovascular, metabolic and socio-economic proof locations for AD are looking between men and lashes. The grasp worth of sex types seems to be sexx with the lookout of sex doing genes, even in mutually related matches 456. PCR job of Y-sequences in Modish cod Arctogadus glacialis and Germany cod Gadus macrocephalus ogac posts that the intention-specific region emerged in anaure more than 7. This could partly be did by work of all-female triploid reports 431To inward determine gender, we time a aspirant PCR kiss including two different wide primers one Y-sequence anti anaturf the other mind a common bar even of the X- and Y- nights and a tinder reverse primer annealing to a anature sex disappointing of the X- and Y-regions selecting in a wide gap in members and every eex males Fig.

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