酿酒酵母新型分泌缺陷突变株突变基因的鉴定 - 南京师范大学图书馆 .pdfVIP

, 2010, 37( 4) : 703708 Journal of Anhui AgriculturalUn ivers ity * 刘泽静, 张 媚, 薛文达, 彭 飞, 许晓风 (, 21004 ) : 对酿酒酵母新型分泌缺陷突变株分泌缺陷特性进行鉴定, 结果发现 Invertase和 Bgl2p的分泌在 37 下受阻, 但 镜观察未见胞内分泌小泡积累用酵母基因组文库挽救突变株, 克隆挽救片段上各候选基因与 pRS315构建重组质粒并转化突变株以检验它们的挽救效果, 发现具有挽救功能的基因是 TEL 2从突变株克隆了 TEL 2的全序列并进行序列测定, 检测到了两 个突变位点, 即: P3 7! L, S545! F对该基因突变前后编码的蛋白质 产物进行了生物信息学分析, 推测出TEL2蛋白可能参与极性外排途径的结构功能域在 24 ~ 434氨基酸范围内 本试验首次揭示了端粒长度调控基因TEL 2在极性外排途径中的重要作用 : 酿酒酵母; 分泌缺陷突变株; TEL 2; 极性外排途径 : Q75 : A : 1 72352X( 2010) 0407030 Characteristics identification of themutated gene in a secretiondeficientm utant strain ofSa ccharomy ces cerevisiae w ith new secretory deficiency perform ance LIU Zejing, ZHANG M ei, XUE W enda, PENG Fei, XU X iaofeng ( College of L ife Sciences, N anjing Norm alUn iversity, Nanjing 21004 ) Abstract: The secretion characters of a new secretiondeficientmutant ofS accharom yces cerev is iae w ere identi fied, and the resu lts show ed both Invertase and Bgl2p secretionw ere blocked at 37 , w h ile no intracellu lar vesicle accumulation w as observed under electron m icroscopy. Yeast genom ic library w as used to transform the mutant to scan the rescue fragmen,t and a rescue fragmentw hich contained tw o genes ofESP 1 andTEL 2w as obtained. ESP 1 andTEL 2w ere cloned separately and ligated to pRS315 to establish the single gene rescue recombinant plasm ids and they w ere transformed into the mutant to test the rescue effect of each cand idate gene. A s a result, only TEL 2 show ed the rescue function. Themutated sequence ofTEL 2w as then cloned from mutant strain and sequenced, and