Screening Of Molecular Marker Of Anhui White Goats Prolificacy Through AFLP Technology

To improve reproductive performance of goats can improve economic benefit.In this study technology of amplified fragment length polymorphism(AFLP) was used in different prolificacy white goats of Anhui province,which can offer basic matetial of Marker Assistant Breeding in prolific Anhui white goats.This experiment designed 8 random primers of EcoRâ… and Taqâ… ,respectively,namely 64 pairs of primer combinations.Twenty individual ear tissue samples were collected from Anhui white goats,ten from high prolificacy,and other from low prolificacy.DNA was extracted from ear tissues and constructed two DNA pools.Technology of AFLP was used to analyze the differences between high and low prolificacy goats,aimed to obtain molecular marker for fecundity of Anhui white goats.The purpose was to improve the Anhui white goats breeding.The result of this study showed the technology of AFLP was suitable for DNA analysis of Anhui white goats.Bands of the AFLP linkage map were clear and abundant,and 32 differential bands were obtained,24 of them were positive bands,8 of them were negative bands.Twelve pairs primers,which amplified clear and abundant bands,were screened from 64 pairs primers that were used in repeated experiment,and obtained two clear differential bands.These two differential bands were extracted through gel extraction kit and purified and cloned and sequenced,and then two sequenceinformations were obtained,one of them was 374bp and the other was 254bp.These two sequences were blasted in NCBI database,the sequence length 374bp had homologous sequences,but the sequence length 254bp had no homologous sequences.If the 374bp sequence was a molecular marker for fecundity of Anhui white goats,a pair primers was designed based on the blast result,and then verified by polymerase chaine reaction restriction fragment length polymorphisms(PCR-RFLP).The result showed two genotypes were found in 104 Anhui white goats,AA genotype were 12,AB genotype were 92 and BB were not found.The frequency of genotype AA in prolific group,low prolificacy group and normal group was 0.9167,0.7123 and 0.9103,respectively.The frequency of genotype AB in three groups was 0.0833,0.2857 and 0.0897,respectively.The frequency of genotype AA in prolific group and normal group were lower than low prolificacy group.It can be inferred that gene B made positive effect in prolificacy in Anhui white goat,and gene A made negative effect in prolificacy.