嗜麦芽黄单胞菌CG样受体跨膜域克隆与分析

根据已知生物LH/CG受体同源性较大的跨膜域序列设计引物，以嗜麦芽黄单胞菌基因组DNA为模板进行PCR扩增，将约600bp目的产物克隆到pUCm-T载体上，经酶切及PCR扩增筛选鉴定得到重组质粒pUCm-Rec，以DIG标记的PCR扩增片段作为探针进行DNA斑点杂交，确证目的PCR扩增片段与该菌染色体DNA有同源性，克隆到的593bpCG样受体跨膜域序列在GenBank中的注册号为AY355346，再以地高辛标记的593bp跨膜域序列为探针，从构建的该菌基因组文库中，筛选到可能与CG样受体属于同一跨膜受体家族的编码组氨酸激酶／效应调节杂合蛋白部分序列的685bp核酸片段(其在GenBank中的注册号为AY359445)。

Cloning and Analyzing of the Transmembrane Region of CG-like Receptor from X.Maltophilia

Using one pair of primers designed by the nucleotide sequence of LH/CG receptor transmembrane region with much homology in known species, and the genomic DNA of X.Maltophilia as template to PCR amplify, and about 600bp aim product was cloned in a pUCm-Tvector. Then pUCm-Rec was screened by restriction endonuclease digestion and PCR identification. The DNA dot hybridization with the DIG-labeled 600bp probe showed that the aim amplified product had homology with the genomic DNA of X.Maltophilia. The cloned 593bp transmembrane region of CG-like receptor was registered for accession number AY355346 on GenBank. 685bp nucleotide sequence(GenBank accession number: AY359445)putatively encoding histidine kinase/response regulator hybrid protein, which maybe belongs to the same transmembrane receptor family with CG-like receptor, was screened by using DIG-labeled 593bp as probe from X.Maltophilia genomic library. This provided clues to study the gene structure and function of CG-like receptor.