【摘要】 根据传染性喉气管炎病毒美国 6 32株和英国Thorne株gC基因的序列设计 1对引物 ,以北京E2株为模板 ,用PCR方法扩增得到长度为 1 9kb的片段 ,并将其克隆至质粒pA T中 .用碱小量法制备质粒DNA ,以酶切和质粒PCR的方法对其进行了鉴定 ,并以正向和反向引物测定其两翼序列 ,结果正向引物测出 498个碱基 ,反向引物测出 499个碱基 ,将所得序列输入GenBank与其他毒株进行比较 ,结果发现其与美国 6 32株的同源性为 94 6 %～ 98 9% ,与英国Thorne株UL44的同源性为 5 5 1% .目前该基因序列已在GenBank中注册 ,代码分别为AF16 10 6 5和AF16 10 6 6 .更多还原

【Abstract】 A pair of primers flanking the gC gene was designed based on published nucleotide sequence of the American 632 strain and British Thorne strain. Expected PCR products of 1.9 kb were obtained after PCR and cloned into pA T vector. The recombinant was designated as pgC and mini preparation of pgC with alkaline protocol and identified with enzyme digestion and plasmid PCR. After sequencing both ends of the gC gene with forward and reverse primer, a 498 bp nucleotide sequence with the forward primer and a 499 bp one with the reverse primer were obtained in the end, their sequences submitted to GenBank and blast searched with other strains. It was found that homology with American 632 strain and British Thorne strain reached 94.6%～98.9% and 55.1% respectively. At present, the gC gene of Beijing E2 strain has been registered with the GenBank, the accession number being AF161065 and AF161066 respectively.更多还原