中文摘要：

根据Genbank数据库已知的链霉菌的查尔酮合成酶基因的保守区设计chs基因特异简并引物，土壤总DNA为模板，利用PCR技术扩增得到该1条chs基因编码区，通过TA克隆、测序和同源比对及进化分析表明：该基因为放线菌来源chs基因．分别在该基因5＇末端和3’末端分别引入的限制酶NcoI和EcoRI酶切位点，利用上述2种限制酶分别酶切导入到psimple—T／chs载体和原核表达pET32a，凝胶回收目的片段后，将二者连接并转化大肠杆菌感受态细胞，转化子经菌液PCR筛选、双酶切鉴定后，3730测序结果表明，该基因全长编码区为1089bp，推测该基因编码全长为362个氨基酸残基，等电点（PI）为5．41、分子量为3965道尔顿含有cHs保守功能区的酸性蛋白质．分析表明，该基因与Streptomyceslividans来源的查尔酮合成酶RppA基因核苷酸相似性高达93％，氨基酸序列相似性高达87．70％．测序结果表明，该基因已经成功插入到pET32a载体中．

英文摘要：

Based on the known conserved cDNA region of streptomycetes chalcone synthase gene in the Genbank database, chs specific primer was designed. Taking soil total DNA as the template, PCR technique was used to get a chs gene coding area. Through TA clone, orderchecking, Homology comparison and evolution analysis, it shows that the gene is genus actinomyces. At the terminal of gene 5 and gene 3 the restriction endonuclease sites of NcoI and EcoR were led in respectively. The two restriction endonuclease digestion were led in the psimple -T/chs carrier and pET32a prokaryotic expression individually. After the Gel extraction of objective fragment, the two were connected and converted into competent Escherichia coli cells. The bacteria liquid was screening by PCR and identified by Double enzyme digestion, and the 3730 order - checking shows that the length of coding region is 1089 bp. It was inferred that the length of the coding region includes 362 amino - acid residue, and the 5.41PI, and 3965 dalton molecular weight has chs conserved functional area acidic protein. The analysis shows that the similarity of nucleotide between the gene and the Chalcone synthase RppA gene of Streptomyces lividans is 93 % , and the similarty of amino acid order is 87.70 %. The order - checking shows that the gene has been successfully inserted into the pET32a carriers.