缺失Cys₄₅-Cys₅₀二硫键的三种突变体凝乳酶原复性的比较研究

缺失Cys 45 -Cys 50 二硫键的三种突变体凝乳酶原复性的比较研究 THE LOCALIZATION OF CHLOROPLAST rbc L GENE AND CONSTRUCTION OF CHLOROPLAST GENE LIBRARY FROM BRASSICACEAE RAPHANUS SATIVA 唐兵 Xiong Yumei 1 first-author 武汉大学生命科学学院 武汉大学生命科学学院 College of Life Sciences, Wuhan University College of Life Sciences, Wuhan University 通过对（Ｃｙｓ→Ａｓｐ）４５／（Ｃｙｓ→Ｓｅｒ）５０、（Ｃｙｓ→Ａｓｐ）４５和（Ｃｙｓ→Ｓｅｒ）５０３种突变体凝乳酶原复性的比较研究，发现：（１）３种突变体凝乳酶原多肽链复性的条件基本一致；（２）ＰＤＩ能有效地促进３种突变体凝乳酶原的复性；（３）３种突变体凝乳酶原的活化条件一致；（４）单突变体凝乳酶原多肽链正确折叠的效率低于双突变体．基于上述结果，我们认为，Ｃｙｓ４５－Ｃｙｓ５０二硫键的形成有利于凝乳酶原多肽链的正确折叠． Improved rapid method of high ionic strength was developed to isolate and purify chloroplast DNA（ctDNA）from Brassicaceae raphanus Sativa. The ctDNA was analyzed with four species restriction endonuclease BamHⅠ,PstⅠ,HindⅢ and EcoRⅠ respectively. Its molecular weight was deduced about 82.5×10 6（125 kb）. Heterologous rbc L gene labeled with Nick Translation as the probe was hybridyzed to ctDNA EcoRI fragment. The result of Southern blot showed that rbc L gene was localized in E11 fragment（1.7 kb）. At last,chloroplast gene library of B.raphanus sativa was constructed via ctDNA EcoRⅠ fragments inserting into vector Bluescript M13 Recombinants and over 15 ctDNA EcoRⅠ fragments were screened. 突变体凝乳酶原 二硫键 复性 Q556.1 国家863高技术项目 1997-04-01 2021-04-01 4