黑曲霉糖化酶基因表达调控的研究──Ⅱ.A.niger T21和3.795glaA基因5'调控区功能的体内分析  

RESEARCH ON THE REGULATION OF GLUCOAMYLASE GENE(glaA) EXPRESSION IN A.NIGER──Ⅱ. ANALYSIS OF THE FUNCTION 0F 5'-REGULATORY REGION OF A. NIGER T21 AND 3.795 glaA GENE

作　　者：乔殿华[1] 钟丽婵[1] 唐国敏[1] 杨开宇[1] 

机构地区：[1]中国科学院微生物研究所,北京100080

出　　处：《微生物学报》1998年第1期26-31,共6页Acta Microbiologica Sinica

基　　金：国家自然科学基金;联合国教科文组织资助

摘　　要：对糖化酶高产菌株A.nigerT21和原始菌株Aniger3．795的glaA5＇上游区的序列分析证明，两者在1．5kb的区域内有9个部位的碱基不同。为考察这些碱基差异是否是引起T21glaA基因转录水平提高的原因，构建了以T21和3．795glaA基因转录调控区及A．nidulans　trpC基因终止子为表达元件的E．colihph基因表达载体（pXH2和pGH1），用pXH2和pGH1分别转化A.nigerT21，对两种转化子的HmB抗性水平测定和Southern杂交分析显示，在转化子XH2C和GH1C中，pXH2和pGH1以相同拷贝数（2拷贝串联）整合到染色体DNA的相同位置上，XH2C的HmB抗性水平（3000μg／ml）为GH1C（1500μg／ml）的2倍。这一结果表明，诱变引起的调控区序列改变使T21glaA基因转录调控区的功能水平比3．795提高1倍。Two plasmid vectors pXH2 and pGH1 were constructed through the fusionof E coli hph gene, the report gene and the 5' upstream regions of A. niger T21and 3.795 respectively, as well as the terminator of A nidulans trpC gene. Theplasmid vectors were than used to transform A niger T21 to functionally identifythose different basic groups between the two 5' upstream regions responsible forhigh-level expression of the glaA gene. Southern analysis of two transformants XH2Cand GH1C revealed that pXH2 and pGH1 were integrated respectively into thechromosome at same site with two copies in tandem array. The level resistant toHmB(3000 μg / ml) of XH2C was twice as high as that (1500 μg / ml) of GH1C,indicating that the changes of basic groups through mutation result in twice increaseof functional level of region responsible for transcription and regulation of A nigerT21 glaA gene compared with that of 3.795.

关 键 词：黑曲霉 糖化酶 基因表达 转录调控区功能 

分 类 号：Q949.327.1[生物学—植物学] TS201.3[轻工技术与工程—食品科学]