WT5BZ] The polymorphic Msp I site in the 5th exon of the bovine growth hormone(bGH)gene was detected by using PCR procedure.A 340bp fragment enclosing the sequence from 2165 position to the 2504 position was amplified...WT5BZ] The polymorphic Msp I site in the 5th exon of the bovine growth hormone(bGH)gene was detected by using PCR procedure.A 340bp fragment enclosing the sequence from 2165 position to the 2504 position was amplified using upper primer 5’GGGCAGATCCTCAAGCAGA 3’ and lower primer 5’ATCCTCCCCCTTGCTGTCC 3’.Genomic DNA samples were from Simmental(n=126),Luxi(n=14),Piemontese(n=8),crossbreed of Piemontese and Nanyang(n=44),Nanyang(n=17),and Hanwoo( Bos taurus koreanae, n=108).PCR products digested with Msp I were separated by electrophoresis in 8% polyacrylamide gel and detected by silver staining.Only a 340bp amplified product could be observed when a Msp I site was absent(UU type).The amplified product,which had a MspⅠ site,showed 245bp and 95bp fragments(CC type).In Simmental and Luxi breeds,only CC was detected.The two polymorphisms(CC and CU)were typed in Piemontese and Nanyang.UU types were observed in Hanwoo breed only.The allele frequencies of C and U were:Piemontese:0.875,0.125;crossbreed of Piemontese and Nanyang:0.818,0 182;Nanyang:0.941,0.059;Hanwoo:0.894,0.106,respectively.PCR products of CC and UU types was sequenced by use of ABI PRISM TM 377 DNA sequencer.A C→T transition was revealed at 2258 position.展开更多
文摘WT5BZ] The polymorphic Msp I site in the 5th exon of the bovine growth hormone(bGH)gene was detected by using PCR procedure.A 340bp fragment enclosing the sequence from 2165 position to the 2504 position was amplified using upper primer 5’GGGCAGATCCTCAAGCAGA 3’ and lower primer 5’ATCCTCCCCCTTGCTGTCC 3’.Genomic DNA samples were from Simmental(n=126),Luxi(n=14),Piemontese(n=8),crossbreed of Piemontese and Nanyang(n=44),Nanyang(n=17),and Hanwoo( Bos taurus koreanae, n=108).PCR products digested with Msp I were separated by electrophoresis in 8% polyacrylamide gel and detected by silver staining.Only a 340bp amplified product could be observed when a Msp I site was absent(UU type).The amplified product,which had a MspⅠ site,showed 245bp and 95bp fragments(CC type).In Simmental and Luxi breeds,only CC was detected.The two polymorphisms(CC and CU)were typed in Piemontese and Nanyang.UU types were observed in Hanwoo breed only.The allele frequencies of C and U were:Piemontese:0.875,0.125;crossbreed of Piemontese and Nanyang:0.818,0 182;Nanyang:0.941,0.059;Hanwoo:0.894,0.106,respectively.PCR products of CC and UU types was sequenced by use of ABI PRISM TM 377 DNA sequencer.A C→T transition was revealed at 2258 position.
