在pFOG405重组质粒中的大肠杆菌碱性磷酸酶基因(PhoA)的启动子及信号肽区的核苷酸顺序被测定并同Kikuchi等人在pYK190重组质粒中所报道的顺序进行了比较。结果指出,包含有PhoA基因启动子及信号肽的PvuⅡ-HpaⅠ片段对于调控外源基因的表达是足够的;启动子5’-端的二元对称结构(26bp)的缺失并不影响外源基因的表达水平。由于pYK190及pFOG405中PhoA基因启动子的上游区核苷酸顺序的不同源性,说明此上游区顺序与PhoA启动子调控下的外源基因表达无关。这一分析为我们利用PvuⅡ-HpaⅡ片段组建高效分泌表达载体及研究基因表达调控提供了有用信息。
The nucleotide sequence of the promoter and the signal peptide of E. coli alkaline phosphotase gene from pFOG405 plasmid has been disclosed and compared with the sequence reported by Kikuchi et al. The result shows that the PvuⅡ-HpaⅡ fragment (144bp) containing the promoter and signal peptide sequence from pYK190 plasmid is sufficient for expression and secretion of foreign genes in E. coli. The deletion of the dyad sequence (26bp) from 5' end of the promoter region doesn't affect expression level of foreign target genes. The experiment also provids the useful information for construction of efficient expression and secretion vector using the PvuⅡ-HpaⅡ fragment and studying regulation of gene expression.
静国忠,刘莉军,蒋美岩.大肠杆菌碱性磷酸酶(PhoA)基因启动子及信号肽核苷酸顺序测定与解析[J].生物化学与生物物理进展,1991,18(6):439-442
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