稻瘟病菌一假定几丁质酶在毕赤酵母中的表达.pdfVIP

() 38 6 Journal of Fujian Agriculture and Forestry Univers ity ( N atural Science Ed ition) 200 11 稻瘟病菌一假定几丁质酶在毕赤酵母中的表达 1 1 1 1 1 2 , , , , , ( 1. 福建农林大学生命科学学院; 2. 福建农林大学植物保护学院, 福建 福州350002) : Guy11 RNA, RTPCR M GG_08054. 6 cDNA , H istag pPIC3. 5K , pPIC3. 5Kw s, GS115, MDMM PCR GS115PICHw s; , . SDSPAGE , 48 ku, ; N iNTA DNS. : ; ; ; + : Q556 . 2 : A : 200 ) Expression of a putativeM agnap orth e ory zae chitinase inP ichia p astoris 1 1 1 1 1 2 ZHOU Jie, CHEN X in , ZHENG X iangzi, LIN Chengzeng , WANG Zonghua , LU Guodong ( 1. College of L ife Science; 2. College of Plant Protection, Fujian Agricu lture and ForestryUniversity, Fuzhou, Fujian 350002, Ch ina) Abstract: Chitinasem ay participate in regulating cellw all dynam ics, m ed iating the grow th and colonization of fungi. In this study, cDNA of a putative chitinase gene (M GG_08054. 6) w as amp lified byRTPCR from the totalRNA ofMagnap orthe oryzae strain Guy 11. It w as then subcloned into theP ichia expression vector pPIC3. 5K w ith a RGSH istag, generating a recomb inant plasm id, pPIC 3. 5Kw s. To express the fusion protein, linearized pPIC3. 5Kw s was transformed into P. pastoris strain GS115 by electroporation. Pos itive recomb inan t clone GS115PICHw s was obtained afterMD p late screen ing and PCR detection. The P ichia cells were in duced bym ethanol to express the secreted recomb inant protein. The relative molecu larw eigh t of recomb inan t protein determ ined by SDSPAGE w as about 48 ku, alm ost equal to the theoretical m olecu lar weight. Recom b inant protein