推定编码嗜麦芽黄单胞菌整合酶类蛋白基因片段的克隆  

Cloning the Gene Fragment Coding the Putative Integrase-like Protein From X Maltophilia

作　　者：梁淑芳[1] 陈曼玲[1] 翟朝阳[1] 

机构地区：[1]四川大学华西基础医学与法医学院生物化学教研室,成都610041

出　　处：《四川大学学报（医学版）》2004年第2期158-160,共3页Journal of Sichuan University(Medical Sciences)

摘　　要：目的　扩增编码嗜麦芽黄单胞菌 CG类受体的核酸序列。方法　根据 Grover报道的嗜麦芽黄单胞菌CG类受体的 342 bp部分核酸序列设计的一特异引物 P1和随机引物进行 PCR扩增 ,将 PCR产物克隆到 p UCm - T载体上 ,对经酶切鉴定筛选的重组质粒上的插入片段进行测序和分析。结果　将约 5 0 0 bp PCR产物克隆到p UCm - T载体上 ,经酶切鉴定筛选得到重组质粒 p U Cm- Int。 p UCm - Int上插入片段经 M13通用引物测序 ,5 10 bp克隆片段 (Gen Bank注册号为 :AY36 396 2 )的 4 10～ 4 86 bp与柑橘黄单胞菌质粒 p XAC6 4上的 XACb0 0 0 9基因的90 34～ 895 8bp部分有 84 %同源序列 ,由 5 10 bp核酸序列编码的 16 9氨基酸 (aa)序列的 4～ 16 6 aa部分与柑橘黄单胞菌 XACb0 0 0 9基因编码的 4 2 4 aa整合酶类蛋白的 38～ 2 0 0 aa序列有 6 2 %同源序列。结论　克隆到的 5 10Objective To amplify the nucleotide sequence of CG-like receptor from X anthomonas maltophilia(X maltophilia). Methods Using the specific primer P1 designed by Grover's reported 342 bp partial nucleotide sequence of X maltophilia CG-like receptor and random primer to PCR amplify, PCR product was cloned in the pUCm-T vector. After the recombinant plasmid was tested by restriction endonuclease digestion, the insert on the recombinant plasmid was sequenced and analyzed. Results About 500 bp PCR product was cloned in the pUCm-T vector and obtained the recombinant pUCm-Int. By sequencing to the insert on the pUCm-Int with M13 universal sequencing primers, the 410-486 bp fragment of the cloned 510 bp nucleotide sequence (GenBank accession number: AY363962) showed 84% identity with the 9304-8958 bp fragment of the XACb0009 gene on plasmid pXAC64 of Xanthomonas axonopodis pv. citri. And the 4-166aa fragment of its translated 169aa sequence had 62% identity with the 38-200 aa sequence of integrase-like protein coded by the XACb0009 gene. Conclusion The cloned 510 bp nucleotide sequence was possibly the partial gene sequence coding the integrase-like protein of X maltophilia.

关 键 词：嗜麦芽黄单胞菌 整合酶类蛋白 随机引物 PCR扩增 

分 类 号：R346[医药卫生—基础医学]