Wang ChangFa

Genetic polymorphism of Nramp1 gene and correlation with mastitis in Holstein cattle

In this research, PCR-SSCP technique was used to analyze the polymorphisms of the exon 11 of Nramp1 gene in Chinese Holstein cattle (n=344), and correlation between polymorphisms of Nramp1 with somatic cell score (SCS) and milk production traits was analyzed. The results show that three genotypes namely AA, BB, and AB were detected. Allele A was predominant and the frequencies of alleles A and B were estimated to be 0.767 and 0.233, respectively. Chi-square test indicated that the polymorphic locus in Chinese Holstein fitted Hardy-Weinberg equilibrium (P>0.05). Sequencing analysis showed two polymorphic sites at positions 200 bp (C/G) and 254 bp (T/G), which resulted in amino acid alteration Ala356Pro and Leu374Met. The least squares means of SCS in Holstein cattle was lower for genotype AA than that for genotypes AB and BB (P<0.05). The least squares means of milk yield of genotype AA and AB were higher than that for genotype BB (P<0.05, P<0.01, respectively). Genotype AA was beneficial to mastitis resistance. This suggested that Nramp1 may be a candidate gene responsible for mastitis in Holstein cattle.

Genetic polymorphism in 5′-flanking region of the lactoferrin gene and its associations with mastitis in Chinese Holstein cows.

Bovine lactoferrin (LF) is a multifunctional glycoprotein found in milk, which acts mainly as a defense factor in the mammary gland. Polymorphism has been found in the bovine LF gene. However, there is no report on genetic polymorphism of LF gene and its associations with mastitis in dairy cattle. In this study, the promoter fragment of LF gene containing -926(G/A), -915(T/G), -478(/G), and +72(T/C) mutations were genotyped by the PCR-RFLP and CRS-PCR method. Two hundred and sixty-eight Chinese Holstein cows were screened. Least square linear model (LSM) analysis was applied to evaluate the associations of LF gene with somatic cell score (SCS). The results indicated that the SCS was significantly affected by -478(/G) and +72(T/C), but not by the other two loci (P >0.05). The SCS of cow with genotype AB in +72(T/C) position was significantly lower than that of genotype AA (P<0.01) or AB (P<0.05). In position -478(/G), the cow with genotype CC showed significantly lower SCS in contrast to cow with genotype CD and DD (P < 0.01). In conclusion, genotype AB in position +72(T/C) and genotype CC in position -478(/G) of LF gene were advantageous genotype, which can be used as candidate markers for mastitis resistance selection in dairy cattle.

CRS-PCR polymorphisms of the GHR gene and its relationship with milk production traits in Chinese Holstein cows

Single-nucleotide polymorphisms (SNPs) of exon 8 of the GHR gene were detected in Chinese Holstein cows by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). The results showed that exon 8 of the GHR gene digested by Tas I could be divided into two kinds of alleles and three kinds of genotypes. The frequencies of allele A and T were 0.6339 and 0.3661, respectively. The frequencies of genotypes AA, AT and TT were 0.459, 0.350 and 0.191, respectively. Sequencing showed one single nucleotide mutation T→A at 4962 bp of the gene in genotype TT when compared with genotype AA, and this mutation resulted in an amino acid change of phenylalanine (TTT)→tyrosine (TAT). The result of χ 2 testing indicated that the genotypic frequency of the GHR gene digested by Tas I did not fit with Hardy–Weinberg equilibrium in this population ( P P