论文部分内容阅读
将乙型脑炎(JE)病毒JaOArS982株的cDNA克隆S22,插入质粒pUC13中,经过一系列加工,构建成pS22×S质粒,用双脱氧链终止法测定cDNA顺序,证明JE病毒RNA5′末端的3000个核苷酸顺序包含了JE病毒结构蛋白的编码。将其中包括V3基因的1600个碱基对的DNA片段切下来插到酵母表达载体YEP133PCT上,构建成pJEV3质粒,经过加工构成pJM105质粒。用其转化啤酒酵母SHY4。在无亮氨酸(leu)的SD琼脂上涂板,分离leu~+表型的转化株,检定V3基因产物的特性。
The cDNA clone S22 of Japanese JE virus strain JaOArS982 was inserted into plasmid pUC13. After a series of processing, a pS22 × S plasmid was constructed and its cDNA sequence was determined by dideoxy chain termination method. The 3000 nucleotide sequence contains the coding for the JE viral structural protein. A DNA fragment of 1600 base pairs including the V3 gene was cut and inserted into the yeast expression vector YEP133PCT to construct a plasmid pJEV3 which was processed to construct the plasmid pJM105. Use it to transform Saccharomyces cerevisiae SHY4. Plates were plated on leu-free SD agar and the leu ~ + phenotype transformants were isolated and the identity of the V3 gene product was determined.