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The Polymorphism Study on the Inhibin Beta Subunit Gene by PCR-RFLP and PCR-SSCP in Small Tail Han Sheep

Author: XueZuo
Tutor: ZhouZhongXiao；ChuMingXing
School: Shanxi Agricultural University
Course: Animal Genetic Breeding and Reproduction
Keywords: Small Tail Han PCR-RFLP PCR-SSCP Inhibiting hormone beta - A subunit gene
CLC: S826
Type: Master's thesis
Year: 2004
Downloads: 161
Quote: 3
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Abstract


This study analyzed the four sheep breeds inhibition of hormone beta-A subunit gene RFLP polymorphism and inhibit hormone beta-B subunit gene SSCP polymorphism. Inhibition of hormone beta-A subunit to the two pairs of primers for PCR-RFLP analysis, the first pair of primers (M1) amplification products 402bp beta-A subunit gene in exon II specific fragment (this study noted INHBA I) No polymorphism was detected. The second pair of primers (M2) amplification product of 1100bp beta-A subunit gene-specific fragment (denoted by INHBA II). Currently no inhibiting hormone beta-A subunit gene sequences reported to inhibit the hormone beta-A subunit gene primers were designed to inhibit hormone beta-A subunit mRNA sequence cattle so by comparing the sheep, the resulting amplified fragment Ⅱ sequence including exon and intron sequences. By restriction enzyme Msp Ⅰ RFLP analysis identified two genotypes, type in a 1.5% agarose gel, digested with respectively 641/183/163/107bp, 641/390/183/163/107bp, denoted as AA and AB. Calculated gene frequencies of these two genotypes within the four sheep breeds, the genotype frequencies of the A gene was dominant allele; inspection certificate gene frequencies and genotype frequencies in Hardy-Weinberg equilibrium. Population genetic characteristics of the INHBA II analysis showed that the gene were tested in four sheep populations in low polymorphism. ANOVA showed, INHBA II genotype Small Tail Han first-born litter size and birth litter size were significantly (P lt; 0.05) The least squares mean significant difference analysis showed that a birth litter size, type AB Small Tail Han prolific than the average AA-0.45 (P lt; 0.05); litter size of the second child, AB Small Tail Han litter size of prolific than the average AA-0.53 (P lt; 0.05). Exon II directly encoding the mature inhibin beta-A subunit, the 402bp fragment sequenced and compared the DNA sequence of a point with the existing nine foreign sheep breeds. Small Tail Han sheep and domestic sheep known sequence diagram completely overlap. In addition to the wild sheep breeds of domestic sheep DNA sequences between the only two base mutation, namely Rhoenschaf sheep 25bp at the G → A cause amino acids expressing mutant Ala → Thr; Merinolandschaf sheep 165 bp at the C → A, did not cause amino acid expressing mutant . Inhibin beta-B subunit gene PCR-SSCP detection, PCR product of 294bp beta-B subunit gene exon II specific fragment polymorphism was not found. By the above results, we believe that the inhibition of hormone beta-subunit A gene can be used as the sheep lambing number of characters molecular genetic markers for marker-assisted selection.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Sheep
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