On the basis of the ovine bone morphogenetic protein 15(BMP15)gene,two pairs of primers(PI and P2)were designed to amplify exons 1 and 2 of the BMP15 gene in five randomly selected does of both Angora and Jining Grey ...On the basis of the ovine bone morphogenetic protein 15(BMP15)gene,two pairs of primers(PI and P2)were designed to amplify exons 1 and 2 of the BMP15 gene in five randomly selected does of both Angora and Jining Grey goats.The sequences of BMP15 exon 1(P1 amplification)of Angora and Jining Grey goats were identical.There was a 3-nucleotide(CTT)insertion in positions 268 to 270 of goat BMP 15 exon1 compared with that of sheep(GenBank accession number AF236078),which caused a leucine insertion in the 12th position of amino acid sequence.Sequence length of goat BMP 15 exon 2(P2 amplification)was identical with that of sheep(AF236079),but there were seven nucleotide and four amino acid changes between goat and sheep.The nucleotide in the 963rd position of BMP15 exon 2 was A for Angora goat and sheep,and G for Jining Grey goat.Based on this A963G mutation,primer pair P3 was designed to detect single nucleotide polymorphism of BMP15 exon 2 in breeds of high prolificacy(Jining Grey),moderate prolificacy(Boer)and low prolificacy(Angora and Inner Mongolia Cashmere)by polymerase chain reactionsingle strand conformation polymorphism(PCR-SSCP).Three genotypes(AA,AG and GG)were detected in Jining Grey goats,two genotypes(AG and GG)in Boer,and only the AA genotype in Angora and Inner Mongolia Cashmere goats.Sequencing revealed one mutation(A963G)in genotype GG compared with genotype AA,and this mutation resulted in an amino acid change of serine→glycine(S300G).In Jining Grey goats,frequencies of AA,AG and GG genotypes were 0.008,0.059 and 0.933,respectively.Genotypic distributions of the BMP 15 gene were significantly different(P<0.05 or P<0.001)between Jining Grey and Boer,Angora,and Inner Mongolia Cashmere goats.In Jining Grey goats,the does with the GG genotype had 0.71(P<0.05)or 1.57(P<0.05)additional kids than did those with AG or AA genotypes,and does with the AG genotype had 0.86(P<0.05)more kids than did those with the AA genotype.These results tentatively indicate that the BMP15 gene is either a major gene that affects prolificacy in Jining Grey goats,or may be a molecular marker in close linkage with such a gene.展开更多
PCR-SSCP was used to detect mutations of bone morphogenetic protein 15(BMP15) gene in both high prolificacy(Small Tail Han sheep,Hu sheep,Jining Grey goat and Boer goat) and low prolificacy breeds(Dorset sheep,Texel s...PCR-SSCP was used to detect mutations of bone morphogenetic protein 15(BMP15) gene in both high prolificacy(Small Tail Han sheep,Hu sheep,Jining Grey goat and Boer goat) and low prolificacy breeds(Dorset sheep,Texel sheep,Inner Mongolia Cashmere goat and Angora goat).Both the nucleotide sequences and the amino acid sequences were compared in amplification fragments of both Small Tail Han sheep and Jining Grey goat.The results indicated that none of the four sheep and the four goat breeds carried the same FecX<sup>R</sup> mutation of the BMP15 gene as do Rasa Aragonesa sheep.The nucleotide sequence of Small Tail Han sheep was completely identical with that of the sheep BMP15 sequence(GenBank AF236079,NM<sub>0</sub>01114767).Three base substitutions(T529G,C530G and T576C) and two amino acid changes(V155G and S171P) were found in Jining Grey goat compared with Small Tail Han sheep.The FecX<sup>R</sup> mutation of the BMP15 gene had no significant effect on high prolificacy of Small Tail Han sheep, Hu sheep,Jining Grey goat and Boer goat.展开更多
基金supported by National Key Basic Research and Development Program of China(No.2006CB102105)National High Technology Research and Development Program of China(No.2006AA10Z139)+1 种基金National Natural Science Foundation of China(No.30540052 and No.30871773)Beijing Natural Sciences Foundation of China(No.6062023)
文摘On the basis of the ovine bone morphogenetic protein 15(BMP15)gene,two pairs of primers(PI and P2)were designed to amplify exons 1 and 2 of the BMP15 gene in five randomly selected does of both Angora and Jining Grey goats.The sequences of BMP15 exon 1(P1 amplification)of Angora and Jining Grey goats were identical.There was a 3-nucleotide(CTT)insertion in positions 268 to 270 of goat BMP 15 exon1 compared with that of sheep(GenBank accession number AF236078),which caused a leucine insertion in the 12th position of amino acid sequence.Sequence length of goat BMP 15 exon 2(P2 amplification)was identical with that of sheep(AF236079),but there were seven nucleotide and four amino acid changes between goat and sheep.The nucleotide in the 963rd position of BMP15 exon 2 was A for Angora goat and sheep,and G for Jining Grey goat.Based on this A963G mutation,primer pair P3 was designed to detect single nucleotide polymorphism of BMP15 exon 2 in breeds of high prolificacy(Jining Grey),moderate prolificacy(Boer)and low prolificacy(Angora and Inner Mongolia Cashmere)by polymerase chain reactionsingle strand conformation polymorphism(PCR-SSCP).Three genotypes(AA,AG and GG)were detected in Jining Grey goats,two genotypes(AG and GG)in Boer,and only the AA genotype in Angora and Inner Mongolia Cashmere goats.Sequencing revealed one mutation(A963G)in genotype GG compared with genotype AA,and this mutation resulted in an amino acid change of serine→glycine(S300G).In Jining Grey goats,frequencies of AA,AG and GG genotypes were 0.008,0.059 and 0.933,respectively.Genotypic distributions of the BMP 15 gene were significantly different(P<0.05 or P<0.001)between Jining Grey and Boer,Angora,and Inner Mongolia Cashmere goats.In Jining Grey goats,the does with the GG genotype had 0.71(P<0.05)or 1.57(P<0.05)additional kids than did those with AG or AA genotypes,and does with the AG genotype had 0.86(P<0.05)more kids than did those with the AA genotype.These results tentatively indicate that the BMP15 gene is either a major gene that affects prolificacy in Jining Grey goats,or may be a molecular marker in close linkage with such a gene.
基金supported by National High Technology Research and Development Program of China(No. 2006AA 10Z 139)National Key Basic Research and Development Program of China(No.2006CB 102105)+3 种基金National Key Technology R&D Program of China(No. 2008BADB2B01,2006BAD01A 11,2006BAD 13B08)National Natural Science Foundation of China(No. 30540052)Beijing Natural Sciences Foundation of China(No.6062023)the special fund for basic scientific research of Institute of Animal Science,Chinese Academy of Agricultural Sciences(No.ywf-rc-1)
文摘PCR-SSCP was used to detect mutations of bone morphogenetic protein 15(BMP15) gene in both high prolificacy(Small Tail Han sheep,Hu sheep,Jining Grey goat and Boer goat) and low prolificacy breeds(Dorset sheep,Texel sheep,Inner Mongolia Cashmere goat and Angora goat).Both the nucleotide sequences and the amino acid sequences were compared in amplification fragments of both Small Tail Han sheep and Jining Grey goat.The results indicated that none of the four sheep and the four goat breeds carried the same FecX<sup>R</sup> mutation of the BMP15 gene as do Rasa Aragonesa sheep.The nucleotide sequence of Small Tail Han sheep was completely identical with that of the sheep BMP15 sequence(GenBank AF236079,NM<sub>0</sub>01114767).Three base substitutions(T529G,C530G and T576C) and two amino acid changes(V155G and S171P) were found in Jining Grey goat compared with Small Tail Han sheep.The FecX<sup>R</sup> mutation of the BMP15 gene had no significant effect on high prolificacy of Small Tail Han sheep, Hu sheep,Jining Grey goat and Boer goat.
