7号淀粉酶链霉菌M1033木糖异构酶基因的启动子活性检测、转录起始点的确定  

Promoter Activity Detection and Transcription Initiation Site Determination of the D-xylose Isomerase Gene of Streptomyces diastaticus strain No.7 M1033

作　　者：杭俊[1] 王玉珍[1] 戴新华 崔涛[1] 牛立文[1] 王淳[1] 徐洵[1] 

机构地区：[1]中国科学技术大学生物系

出　　处：《Acta Genetica Sinica》1995年第3期239-244,共6页

基　　金：国家高技术"863"项目

摘　　要：分别从重组质粒ｐＵＢ１及其Ｍ（１３）亚克隆Ｓ１中将７号淀粉酶链霉菌（ＳｔｒｅｐｔｏｍｙｃｅｓｄｉａｓｔａｔｉｃｕｓＮｏ．７）Ｍ１０３３（以下简称Ｓ．ｄｉ．Ｍ１０３３）木糖异构酶基因的－１９２－＋５８１ｂｐ片段克隆入链霉菌启动子探测质粒ｐＩＪ４０８３中，转化变铅青链霉菌（Ｓ．ｌｉｖｉｄａｎｓ）ＴＫ２４。通过对其邻苯二酚加双氧酶活性的检测表明，该片段具有启动子活性。应用Ｍ１３亚克隆Ｓ１和合成引物Ｐ６延伸制备放射性标记的单链ＤＮＡ探针；通过Ｓ．ｄｉ．Ｍ１０３３的总ＲＮＡ的Ｓ１核酸酶保护实验，确定了其转录的起始位点，并由此探讨了与木糖异构酶基因表达有关的一些因素。The DNA segment of the Streptomyces diastaticus strain No.7 M1033 D-xylose Isomerase gene,from -192 to +581 bp according to the translation start site(tss).was cloned in Streptomyces promoter-probe plasmid pIJ4083 from recombination plasmid pUB1 and its M13 subclone S1 respectively.The protoplasts of S.Iividans TK24 were transformed with the recominant,and then detected the promoter activity by measuring the catethol dioxygenase expression.Results of S1 mapping of total RNA of S.di.M1033 indicatedthat:the transcription initiation site of D-Ⅺ gene was 40bp upstream the coding region;there was another gene encoded on the antisense strand upstream D-Ⅺ gene,with a 114-nucleotide sequence separating their coding regions.According to its DNA sequence,we believed it was the xylulose kinase gene Data from quantitive S1 mapping also suggested that transcription of the two genes was induced by D-xylose and repressed by glucose.

关 键 词：启动子 转录起始位点 链霉点 木槽异构酶基因 

分 类 号：Q939.103[生物学—微生物学]