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ABSTRACT: In this study, we cloned the coding sequence of chicken CRBP IV, quantified the mRNA expression in Erlang Mountainous Chickens, and investigated a polymorphism in this gene and its association with egg production traits among 349 individuals. The cloned fragment contained a 384 bp open reading frame, which encoded a predicted protein of 127 amino acids and was highly conserved among species. Expression of CRBP IV mRNA was detected in all eight tissues (small intestine, heart, liver, kidney, oviduct, ovary, pituitary, and hypothalamus) at different ages (12, 24, 32 and 45 w). High expression was found in small intestine, pituitary, kidney and liver, whereas it was low in the heart (p < 0.05). The CRBP IV mRNA levels changed with age in the various tissues, and were highly expressed in all tissues at 32 w, except for the heart. We identified one nucleotide substitution (c. 826T>C) in the second exon, which caused an amino acid change (p. S49L). Genotypes (TT, TC and CC) had significant effects on the age at first egg (AFE), total eggs for 300 days (TE300) and highest continuous laying days (HCLD). The CC genotype would be genetically advantageous to improve egg production traits due to earlier AFE, more TE300, and longer HCLD.
International Journal of Molecular Sciences 01/2013; 14(3):4432-43. · 2.60 Impact Factor
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ABSTRACT: The Mustang, Musculoskeletal Temporally Activated Novel-1 Gene (MUSTN1) plays an important role in regulating musculoskeletal development in mammals. We evaluated the developmental and tissue-specific regulation of MUSTN1 mRNA and protein abundance in Erlang Mountainous (EM) chickens. Results indicated that MUSTN1 mRNA/protein was expressed in most tissues with especially high expression in heart and skeletal muscle. The MUSTN1 protein localized to the nucleus in myocardium and skeletal muscle fibers. There were significant differences in mRNA and protein abundance among tissues, ages and between males and females. In conclusion, MUSTN1 was expressed the greatest in skeletal muscle where it localized to the nucleus. Thus, in chickens MUSTN1 may play a vital role in muscle development.
International Journal of Molecular Sciences 01/2013; 14(3):5545-59. · 2.60 Impact Factor
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ABSTRACT: Because wild rhesus macaque (Macaca mulatta) populations have suffered major declines, there is a growing need to characterize their genetic and population structure in order to protect the genetic integrity of this species. In this study, we genotyped a sample comprising 120 wild rhesus macaques from six sites in Sichuan Province for 30 nuclear microsatellite (STR) loci using an ABI 3130xl genetic analyzer. Bayesian analyses and PCA clearly differentiated monkeys from Heishui from those at other sites. The samples from all six sites exhibited high gene diversity suggesting that the Sichuan wild rhesus macaque populations are not threatened by a lack of genetic diversity. Deviation from Hardy-Weinberg equilibrium was more frequent in the Danba and Heishui populations. This may be due to the more fragmented habitat and less disturbance by humans in this area that foster greater subpopulation structuring than occurs in eastern China. We suggest that this population subdivision is the result of both long-term geographic barriers and human activity.
Molecular Biology Reports 12/2012; · 2.93 Impact Factor
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ABSTRACT: POU1F1 is an essential factor that regulates the development and reproduction of animal. The objective of the current research was to screen for polymorphism, expression of POU1F1 and their association with carcass quality traits. A total of 126 Erlang mountainous chickens from two strains (SD02 and SD03) were employed for testing. Seventeen single nucleotide polymorphisms (SNPs) were detected, but only two SNPs (g.96217999 T > C and g.96219442 C > T) were associated with carcass quality traits. In SD03 chicken, g.96217999 T > C genotypes were significantly associated with body weight (BW), carcass weight (CW), eviscerated weight (EW), and semi-eviscerated weight (SEW; P < 0.05), and was highly significantly associated with breast muscle weight (BMW) and abdominal fat weight (AW; P < 0.01). g.96219442 C > T was significantly associated with BW, EW, SEW (P < 0.05). However, these two SNPs were not significantly associated with any carcass traits in SD02 chicken. Diplotypes showed that in SD03 chicken, the haplotype [C: C] was the most favorable haplotype because it was associated with higher BW, CW, SEW, EW, BMW, and AW (P < 0.05). On the contrary, haplotype [T: T] was associated with lower carcass quality traits (P < 0.01). In addition, qRT-PCR revealed that at 13 weeks, the POU1F1 mRNA expression was significantly higher in breast muscle of cock compared to that of hens (P < 0.05), whereas there was no significant correlation between POU1F1 expression and carcass traits. These results suggested that POU1F1 could be a potential candidate gene for carcass traits in chicken.
Molecular Biology Reports 06/2012; 39(8):8363-71. · 2.93 Impact Factor
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ABSTRACT: In the current research, the polymorphism of FATP4 gene was analyzed in Erlang Mountainous chickens. A total of nine genetic variants were identified by FATP4 gene sequencing analysis across the chicken samples. Significant associations (p < 0.05) were observed for two SNPs (g.5608778C>T and g.5608814G>A in exon 6) with certain carcass traits (such as live weight, carcass weight, eviscerated weight) in S01 and S05 populations, respectively. Meanwhile, in S05 population, haplotype 3 (T-G) and haplotype 2 (C-A) were associated with higher and lower partial carcass traits such as live weight, carcass weight, eviscerated weight and semi-eviscerated weight, respectively. Moreover, we investigated the expression profile of this gene during ontogenesis in Mountainous black-boned chicken. Quantitative real-time PCR analysis showed that FATP4 mRNA had the highest expression level in small intestine tissue over all other tissues examined. The FATP4 mRNA levels presented remarkable developmental changes with age in the various tissues. These results suggested that the FATP4 gene might play an important role in controlling chicken carcass traits.
International Journal of Molecular Sciences 01/2012; 13(6):6820-35. · 2.60 Impact Factor
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ABSTRACT: Calpain-3 is a skeletal muscle-specific protease and participates in the regulation of myogenesis. In this study, we quantified the expression of calpain-3 (CAPN3) mRNA in a Chinese local chicken breed (Sichuan Mountainous Black-boned chicken [MB]), to discern the tissue and ontogenic expression pattern. Meanwhile, we compared the CAPN3 mRNA expression pattern in MB chicken at 10 weeks with a commercial meat type chicken line (S01) of the same age to identify the unique expression pattern under different genetic background. A real time quantitative PCR (qRT-PCR) assay was developed for an accurate measurement of its expression in various tissues from chickens at different ages (0, 2, 4, 6, 8, 10, and 12 weeks). Expression of the CAPN3 mRNA was detected in the selected tissues, regardless of age. The breast muscle and leg muscle tissues had a significantly higher expression than the other tissues from the same individual (P < 0.01). Overall, the CAPN3 mRNA level exhibited a "rise-decline" developmental change in detected tissues except for brain. The S01 chicken had a higher expression of the CAPN3 mRNA in detected tissues than the MB chicken at 10 weeks. The present expression data of chicken CAPN3 gene may provide some information to shed light on the tissue and ontogenic expression pattern during chicken development.
Molecular Biology Reports 06/2011; 39(4):3517-21. · 2.93 Impact Factor
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ABSTRACT: The calpain system, a Ca(2+)-activated protease family, plays an important role in postmortem tenderization of skeletal muscle due to its involvement in the degradation of important myofibrillar and associated proteins, as well as in cytoskeletal remodeling and regulation of muscle growth. In this study, we quantified the expression of calpastatin (CAST) in two Chinese chicken breeds (mountainous black-bone chicken breed (MB) and a commercial meat type chicken breed (S01)), to discern the tissue and age-related specific expression pattern and its potential role on muscle tissue metabolism. Real-time quantitative PCR (RT-qPCR) assay was developed for accurate measurement of CAST mRNA levels in various tissues from chicken with different ages (0, 2, 4, 6, 8, 10, and 12 week). CAST mRNA was detected in collected organs. The heart and leg muscle tissues had the highest expression of CAST than other tissues from the same chicken (P < 0.01). Age-related expression pattern of CAST gene was evident in breast muscle, liver, and brain tissues (P < 0.05), but not in heart and leg muscle tissues (P > 0.05). Overall, the CAST mRNA level exhibited a "rise-decline-rise-decline" developmental change in breast muscle and liver, with the highest expression at 2 weeks and the lowest expression at 8 weeks. The S01 chicken had significantly higher expression of CAST in breast muscle and heart than the MB chicken (P < 0.05) at 10 weeks. Our results suggested the CAST expression may be related to muscle fiber development.
Molecular Biology Reports 06/2011; 39(2):1839-43. · 2.93 Impact Factor
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ABSTRACT: Between one and six subspecies of Chinese rhesus macaques (Macaca mulatta) have been proposed based on morphological differences and/or their geographic distribution. In this study, a 489 base pair fragment of the mitochondrial control region was amplified from 230 DNA samples collected from rhesus macaques in the Sichuan province in Western China. The fragment was then sequenced and aligned with 208 sequences from wild rhesus macaques, sampled throughout the species' geographic range in China downloaded from GenBank. Phylogenetic analysis of the 182 unique sequences identified among these samples divided Chinese rhesus macaques into two western haplogroups (haplogroups A and B) and three older eastern haplogroups (haplogroups C, D, and E), whose differentiation probably occurred during the penultimate glacial event. During the warming after the penultimate glacial event, haplogroups A, B, and E rapidly expanded and a relatively young subhaplogroup of haplogroup E, E', limited to Southern China but shared with Vietnamese rhesus macaques, was reintroduced from Indochina during the last glacial event. One haplotype most closely related to subhaplogroup E' probably represents the isolation of Hainan Island, to where it is restricted, from the mainland by the formation of the Qiongzhou Strait approximately 8,500 years ago. The distribution of haplogroups both informs the phylogeographic history of dispersal of Chinese rhesus macaques and has implications for their suitability as animal models in biomedical research.
American Journal of Primatology 04/2011; 73(9):883-95. · 2.22 Impact Factor
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JSW. 01/2011; 6:1337-1344.
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ABSTRACT: The solute carrier family 27 (SLC27, also known as fatty acid transport proteins [FATPs]) plays important biological roles in cells. However, there is no report about the expression profile of SLC27 member in chicken. In this study, we quantified the expression of SLC27A1 (FATP1) mRNA in a mountainous black-boned chicken breed (MB) and a commercial meat type chicken breed (S01), to discern the tissue and age-related specific expression pattern and their potential involvement in fat deposition and muscle fatty acid metabolism. Real-time quantitative PCR assays were developed for accurate measurement of SLC27A1 mRNA levels in different tissues from chicken with different ages (0-12 weeks). Expression of SLC27A1 mRNA was detected in all tissues examined. There was a significantly age-related change of the SLC27A1 mRNAs in heart, breast muscle (BMW), leg muscle (LMW), liver, and abdominal fat (AF) tissues (P < 0.05). The breast muscle and leg muscle tissues had the highest expression of SLC27A1 mRNA than the other tissues from the same individual at 0, 2 and 4 weeks. The overall SLC27A1 mRNA level exhibited a "rise-decline" developmental change in all tissues except for breast muscle, subcutaneous fat, and brain. The S01 chicken had a higher expression of the SLC27A1 mRNA in breast muscle, subcutaneous fat, and heart tissues than the MB chicken. Our results showed that the expression of SLC27A1 mRNA in chicken tissues exhibits specific developmental changes and age-related patterns.
Molecular Biology Reports 12/2010; 38(8):5139-45. · 2.93 Impact Factor
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ABSTRACT: In this study, we aimed to detect the single nucleotide polymorphisms (SNPs) of the chicken FATP1 gene and discern the potential association between FATP1 SNPs and chicken carcass traits. A total of 620 meat-type quality chickens from six commercial pure lines (S01, S02, S03, S05, S06 and D99) and two cross lines (S05 × S01 and S06 × S01) were screened by using the single-strand conformational polymorphism analysis (SSCP) and DNA sequencing. Five SNPs [g.49360G > A, g.48195G > A, g.46847A > G, g.46818A > G, and g.46555A > G] were identified in chicken FATP1 gene. SNP g.46818 A > G was a rare variant and was not considered in the subsequent analysis. Sixteen haplotypes were reconstructed on the basis of the other four SNPs. The linear regression model analysis indicated that there were significant associations of certain diplotypes with part of carcass traits, such as live weight (LW), carcass weight (CW), and semi-eviscerated weight (SEW) (P < 0.05). In particular, diplotype H2H4 had a negative effect on LW, CW, SEW, and abdominal fat weight (AW); diplotype H6H10 had the highest reducing effect on subcutaneous fat thickness (SFT). Our results suggested that FATP1 gene polymorphisms were associated with chicken carcass traits or was linked with the major gene. The SNPs in this gene may be utilized as potential markers for marker-assisted selection (MAS) during chicken breeding.
Molecular Biology Reports 03/2010; 37(8):3683-90. · 2.93 Impact Factor
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ABSTRACT: We quantified chicken calpain 2 (CAPN2) expression in two Chinese chicken breeds (mountainous black-bone chicken breed [MB] and a commercial meat type chicken breed [S01]) to discern the tissue and ontogenic expression pattern and its effect on muscle metabolism. Real-time quantitative PCR assay was developed for accurate measurement of the CAPN2 mRNA expression in various tissues from chickens of different ages (0, 2, 4, 6, 8, 10, and 12 weeks). Results showed that the breast muscle and leg muscle tissues had the highest expression of CAPN2 compared to the other tissues from the same individual (P < .05). Overall, the CAPN2 mRNA level exhibited a "rise" developmental change in all tissues. The S01 chicken had a higher expression of the CAPN2 mRNA in all tissues than the MB chicken. Our results suggest that chicken CAPN2 expression may be related to chicken breeds and tissues.
Molecular biology international. 01/2010; 2010:373241.
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ABSTRACT: Recognition of natural bitter toxins through taste is one of the most effective mechanisms of self-safety. An approximate 1 169 bp sequence of the bitter taste receptor T2R2 gene was obtained by PCR and cloning technique from hog badger genomic DNA(GenBank accession number: FJ812727). This sequence contains a complete single exon (without intron) 915 bp in size, which encodes 304 amino acid residues. The isoelectric point (pI) of the protein is 9.76 and its molecular weight is 34.74 kDa. Topology prediction showed that the T2R2 protein contained one N-glycosylation site, one N-myristoylation site, and two potential protein kinase C phosphorylation sites. Additionally, the whole peptide chain was comprised of seven transmembrane helix regions, four extracellular regions, and four intracellular regions. The T2R2 is a hydrophobic protein with a few hydrophilic components. Homology analysis of the T2R2 gene sequences by Clustal W indicated that the cDNA sequence homology of T2R2 gene in hog badger with dog, cat, cattle, horse, chimpanzee, and mouse is 91.4%, 90.6%, 84.4%, 85.4%, 83.8%and 72.1%, respectively, and the homology of amino acid sequence is 85.5%, 85.8%, 74.0%, 77.6%, 75.3% and 61.5%, respectively. The results of nucleotide acid substitution computation and selective test showed that strong purifying selection (functional constraint) occurred between hog badger and the six species, respectively, which mainly existed in the transmembrane regions of T2R2. In addition, the Neighbour-Joining tree of T2R2 gene exons from these seven species is consistent with their species tree, indicating that the T2R2 gene is suitable for constructing molecular phylogenetic tree among different species likewise.
Hereditas (Beijing) 11/2009; 31(11):1113-20.
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ABSTRACT: Adipose differentiation-related protein gene (ADP) plays an important role in controlling lipid accumulation in mammals. It may also affect lipid deposition in birds. However, the molecular mechanism of its actions in birds remains unknown. In the present study, the coding sequence of ADP cDNA for Chinese native breed Sichuan Mountainous Black-bone chicken (MB) was first cloned from abdominal fat using reverse transcription-PCR (RT-PCR). This putative MB ADP cDNA (1,881 bp) encodes an open reading frame of 438 amino acids (AA) and shares high AA sequence identity with that of red jungle fowl (99%), duck (92%), house mouse (70%), human (70%), chimpanzee (70%), pig (70%), domestic cow (69%) and domestic sheep (68%). Further analysis using bioinformatics shows the deduced MB ADP protein has the typical characters of PAT (Perilipin, Adipophilin and Tip47) family. Quantitative real-time PCR (qRT-PCR) analysis revealed that ADP expresses in chicken leg muscle, whole brain, heart, liver, pectoralis muscle, abdominal fat and subcutaneous fat. Ontogenetic expression studies shows ADP expression levels in abdominal fat, subcutaneous fat and pectoralis muscle were prior to that in leg muscles at posthatch day (P) 84. But, its levels in abdominal fat and subcutaneous fat were less than that in leg muscles at P28, 42, 56 and 70, respectively. The ADP expression levels in subcutaneous fat and abdominal fat were stable from P28 to P70 and both were less than their counterparts at P84. However, it changed greatly in pectoralis muscle, liver, brain, heart and leg muscle at all points. Patterns of ADP expression suggest that ADP plays an important role in fat development, but further study is needed to reconfirm its function in a large population and in other breeds with different genetic backgrounds.
Molecular Biology Reports 09/2009; 37(6):2839-45. · 2.93 Impact Factor
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ABSTRACT: Myocyte-specific enhancer-binding factor 2A (MEF2A) gene is a member of the myocyte-specific enhancer-binding factor 2 (MEF2) protein family which involved in vertebrate skeletal muscle development and differentiation. The aim of the current study is to investigate the potential associations between MEF2A gene SNPs (single nucleotide polymorphisms) and the carcass traits in 471 chicken samples from four populations. Three new SNPs (T46023C, A72626G, and T89232G) were detected in the chicken MEF2A gene. The T46023C genotypes were associated with live body weight (BW), carcass weight (CW), eviscerated weight, semi-eviscerated weight (SEW), and leg muscle weight (LMW) (P < 0.05); the A72626G genotypes were associated with BW, CW, LMW (P < 0.01) and breast muscle weight (BMW), leg muscle percentage (LMP) (P < 0.05); whereas the T89232G genotypes were associated with carcass percentage (CP) and semi-eviscerated percentage (SEP) (P < 0.05). The haplotypes constructed on the three SNPs were associated with BW, CW, LMW (P < 0.01), SEW, BMW, CP (P < 0.05). Significantly and suggestive dominant effects of diplotype H1H2 were observed for BW, CW, SEW, BMW and CP, whereas diplotype H5H5 had a negative effect on BW, CW, SEW, BMW and LMW. Our results suggest that the MEF2A gene may be a potential marker affecting the muscle trait of chickens.
Molecular Biology Reports 09/2009; 37(1):587-94. · 2.93 Impact Factor
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ABSTRACT: Using PCR-SSCP with five primer pairs, we detected six single nucleotide polymorphisms of the H-FABP gene: 332G --> A, 534G --> A, 783C --> T, 835C --> T, 1198T --> C, and 2329C --> T. Chi-square results showed significant differences (P < 0.05) in genotype frequency among breeds in Fragment 1 and extremely significant differences (P < 0.01) in Fragments 2-4. We found a significant association between Fragment 2 genotype and muscle fiber number, Arg and Thr (P < 0.05); between Fragment 3 genotype and living weight, carcass weight, breast muscle weight, abdominal fat weight, and abdominal fat percentage (P < 0.05); between Fragment 4 genotype and Thr, Phe, and inosinic acid (P < 0.05). It was concluded that H-FABP was the major gene influencing slaughter performance and meat quality or was linked with the major gene in these strains and that the C783T mutation could be used as a candidate molecular genetic marker for breeding selection. The combination M1C2-B2B2-D1D1 is an ideal model for breeding in these strains because it can improve slaughter and meat quality traits.
Biochemical Genetics 06/2009; 47(7-8):511-20. · 0.86 Impact Factor
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ABSTRACT: Melanocortin 1 receptor (MC1R) gene plays a key role in determining coat color in several species, including the cattle. However, up to now there is no report regarding the MC1R gene and the potential association of its mutations with coat colors in yak (Poephagus grunniens). In this study, we sequenced the encoding region of the MC1R gene in three yak breeds with completely white (Tianzhu breed) or black coat color (Jiulong and Maiwa breeds). The predicted coding region of the yak MC1R gene resulted of 954 bp, the same to that of the wild-type cattle sequence, with >99% identity. None of the mutation events reported in cattle was found. Comparing the yak obtained sequences, five nucleotide substitutions were detected, which defined three haplotypes (E(Y1), E(Y2), and E(Y3)). Of the five mutations, two, characterizing the E(Y1) haplotype, were nonsynonymous substitutions (c.340C>A and c.871G>A) causing amino acid changes located in the first extracellular loop (p.Q114K) and in the seventh transmembrane region (p.A291T). In silico prediction might indicate a functional effect of the latter substitution. However, all three haplotypes were present in the three yak breeds with relatively consistent frequency distribution, despite of their distinguished coat colors, which suggested that there was no across-breed association between haplotypes or genotypes and black/white phenotypes, at least in the investigated breeds. Other genes may be involved in affecting coat color in the analyzed yaks.
Journal of Biomedicine and Biotechnology 02/2009; 2009:861046. · 2.44 Impact Factor
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ABSTRACT: The aim of this study is to screen single nucleotide polymorphisms (SNP) of chicken Calpain3 (CAPN3) gene and to analyze the potential association between CAPN3 gene polymorphisms and carcass traits in chickens. We screened CAPN3 single nucleotide polymorphisms (SNP) in 307 meat-type quality chicken from 5 commercial pure lines (S01, S02, S03, S05, and D99) and 4 native breeds from Guangdong Province (Huiyang Huxu chicken and Qingyuan Ma chicken) and Sichuan Province (Caoke chicken and Shandi Black-bone chicken), China.
Two SNPs (11818T>A and 12814T>G) were detected by single strand conformation polymorphism (SSCP) method and were verified by DNA sequencing. Association analysis showed that the 12814T>G genotypes were significantly associated with body weight (BW), carcass weight (CW), breast muscle weight (BMW), and leg muscle weight (LMW). Haplotypes constructed on the two SNPs (H1, TG; H2, TT; H3, AG; and H4, AT) were associated with BW, CW (P < 0.05), eviscerated percentage (EP), semi-eviscerated percentage (SEP), breast muscle percentage (BMP), and leg muscle percentage (LMP) (P < 0.01). Diplotype H1H2 was dominant for BW, CW, and LMP, and H2H2 was dominant for EP, SEP, and BMP.
We speculated that the CAPN3 gene was a major gene affecting chicken muscle growth and carcass traits or it was linked with the major gene(s). Diplotypes H1H2 and H2H2 might be advantageous for carcass traits.
BMC Genetics 01/2009; 10:10. · 2.47 Impact Factor
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ABSTRACT: SSCP (Single-strand conformation polymorphism, SSCP) technique was applied in this study to analyze the population genetic information about genetic distribution, variation and heterozygosity of PRKAB2(Protein kinase, AMP-activated, beta 2 non-catalytic subunit) gene in five purelines and three crossbreds. The genetic effects of PRKAB2 gene on fiber density and partial carcass traits were also analyzed. Only one point mutation T-->C at base position 406 located among the first exon of the whole CDS was found in the whole CDS.The wild-type allele A had significant positive effects on live-weight, carcass-weight, leg-muscle-weight and abdomen-fat-weight (P<0.05), and had positive effects on meat tenderness.
Hereditas (Beijing) 08/2008; 30(8):1033-8.
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ABSTRACT: In order to obtain the stable PCR combinations for silkies genetic analysis, multiplex PCR was used and its reaction condition was optimized. Five combinations of multiplex PCR with good effects were obtained from eighteen microsatellite markers. Three of them were quadruplex-PCRs, and two were triplex-PCRs. The products of multiplex PCR were further combined into three sets for the electrophoresis on ABI 3100-Avant Genetic Analyzer. The results suggested that the eighteen microsatellite markers could be successfully applied to the silkies genetic analysis.
Hereditas (Beijing) 05/2008; 30(4):521-6.
