家蝇金属硫蛋白基因的克隆_原核表达和活性检测.pdfVIP

Acta Entomolog ica Sinica, April 2010, 53( 4): 379 - 384 ISSN 张 迪, 任国栋, 唐 婷, 董晓寅, 柳峰松* (, 071002) : , EST , RACE 1M usca domestica M dM tn ( GenBank GU289398) , M dM tn cDNA 408 bp, 1 123 bp, 40, 10, CXC 38 kD, 878 , pETDsbAMT , Escherichia coli BL21( DE3) MT , MdM tn : ; ; ; ; : Q966 : A : 20 10) Cloning, prokaryotic expression and activity detection of the metallothionein gene inM usca dom estica (Diptera: Muscidae) * ZHANG D,i REN GuoDong, TANG T ing, DONG X iaoY in, LIU FengSong ( College of L ife Sciences, HebeiUn iversit, Baoding, H ebei 071002, Ch ina) Abstract: M etallothioneins (MTs) are low molecular w eigh,t metalb ind ing and c steine rich proteins found in a variet of living organism s In th is stud , a 408 bp cDNA encod ing for a m etalloth ionein w as cloned from housefl (M usca dom estica ) b RACE based on EST information and nam ed as MdM tn ( GenBank accession no GU 289398) Sequence anal sis show ed that MdM tn contains a 123 bp open read ing frame (ORF) encod ing a protein of 40 am ino acid residuesTheM dM tn peptide sequence includes 10 c steine residues w ith a d istribution pattern of CXCThe predicted molecular w eight of encoding protein is 38 kD w ith the isoelectric point ( pI) of 878 In order to detect theM dM tn activit in binding heav m etals, the target gene w as cloned into a prokar otic expression vector pETD sbA, and then a fusion proteinw as expressed in Escherichia coli BL21( DE3) It w as found that in the presence of CdCl, the 2 2+ expression ofM dM tn significantl in