推定编码嗜麦芽黄单胞菌整合酶类蛋白基因片段的克隆

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pUCm-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500 bp PCR产物克隆到pUCm-T载体上，经酶切鉴定筛选得到重组质粒pUCm-Int。pUCm-Int上插入片段经M13通用引物测序，510 bp克隆片段(GenBank注册号为：AY363962)的410～486 bp与柑橘黄单胞菌质粒pXAC64上的XACb0009基因的9034～8958 bp部分有84％同源序列，由510 bp核酸序列编码的169氨基酸(aa)序列的4～166aa部分与柑橘黄单胞菌XACb0009基因编码的424aa整合酶类蛋白的38～200aa序列有62％同源序列。结论克隆到的510 bp核酸序列可能是编码嗜麦芽黄单胞菌整合酶类蛋白的部分基因序列。

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

Objective 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.