几丁质酶表达质粒pKChiA和pMChiA的构建及表达  

CONSTRUCTION OF THE PLASMID pKChiA AND pMchiA CARRYING CHITINASE GENE AND THEIR EXPRESSION IN E.coli

作　　者：叶志华[1] 梁建庆[1] 

机构地区：[1]广东省微生物研究所广东省菌种保藏选育重点实验室,广州510070

出　　处：《微生物学通报》2001年第6期35-39,共5页Microbiology China

基　　金：广东省自然科学基金资助项目 (No . 970 6 88)

摘　　要：从质粒pLCHIA中切出含粘质沙雷氏菌 (Serratiamarcescens)几丁质酶基因 (chiA)的 1 8kbHinfI片段 ,将其插入到表达载体pKK2 2 3 3强启动子Ptac下游的SmaI位点内 ,构建成几丁质酶表达质粒pKChiA。再用内切酶BamHI从pKChiA质粒中切出 2 1kbPtac ChiA片段 ,并将其重组到质粒pMC71A的单一BamHI位点内 ,构建成另一种几丁质酶表达质粒pMChiA。这两种质粒可在大肠杆菌HB1 0 1和JM1 0 5中高效表达 ,几丁质酶表达水平比质粒pLCHIA高 1～ 3倍。A 1.8 kb HinfI fragment carrying the chitinase gene(chiA) from Serratia marcescens was isolated from the plasmid pLCHIA and was inserted into the SmaI site downstream of the strong Ptac promoter in the expression vector pKK223-3,yielding the plasmid pKChiA.The 2.1 kb PtacChiA fusion fragment was excised by BamHI from the plasmid pKChiA and was inserted into the single BamHI site present within the plasmid pMC71A,thus generating the plasmid pMChiA.High levels of chitinase were produced by the E.coli strains HB101 and JM105 carrying the plasmid pKChiA or the plasmid pMChiA,with the amount of production were higher 1～3 fold than that produced by the strains carrying the plasmid pLCHIA.

关 键 词：几丁质酶基因 pKChiA质粒 pMChiA质粒 粘质沙雷氏菌 表达 

分 类 号：Q782[生物学—分子生物学]