三株烟草天蛾新细胞系的生长特性及重组蛋白表达  

Growth characteristics and expression of recombinant proteins in three new cell lines from Manduca sexta(Lepidoptera:Sphingidae)

作　　者：姜磊[1] 李国勋[1] 李长友[1] Robert R.GRANADOS Gary W.BLISSARD 

机构地区：[1]青岛农业大学中美无脊椎动物细胞工程中心,山东青岛266109 [2]Boyee Thompson Institute for Plant Research(BTI)at Cornell University,Ithaca,NY 14853,U.S.A

出　　处：《昆虫学报》2010年第11期1227-1232,共6页Acta Entomologica Sinica

基　　金：国家重点基础研究发展规划(“973”计划)项目(2009CB118902);国家自然科学基金项目(30771451)

摘　　要：从尚未涉及的昆虫种类中建立新的细胞系能为基础研究和生物技术应用提供重要资源。本实验通过细胞培养技术,建立了3株来源于鳞翅目昆虫烟草天蛾Manduca sexta卵组织的新细胞系,分别命名为QB-Ms1-8,QB-Ms2-2和QB-Ms2-7。这3株细胞已经培养在TNM-FH培养基中,28℃条件下传代培养了约50代,大部分细胞呈梭形,细胞群体倍增时间分别为51,31和49h。虽然这3株细胞系对苜蓿银纹夜蛾核型多角体病毒(Autographa californica multiple nuclear polyhedrosis virus,AcMNPV)不够敏感,侵染后96h感染率在33%～40%之间,但是QB-Ms2-2细胞与BTI-Tn5B1-4细胞比较,分泌型碱性磷酸酶(SEAP)活性表达更高。本研究从建立的3株烟草天蛾新细胞系中筛选出SEAP高表达的细胞系QB-Ms2-2,为进一步细胞克隆和筛选提供了新资源。The development of new cell lines from unusual and non-pest species may provide important source for basic research and biotechnology application.In this study,three new insect cell lines designated as QB-Ms1-8,QB-Ms2-2,and QB-Ms2-7,respectively,were established from eggs of Manduca sexta.These cells were subcultured approximately 50 passages in TNM-FH medium at 28℃.The majority of cells of the lines generally are spindle-shaped.The population doubling time of QB-Ms1-8,QB-Ms2-2 and QB-Ms2-7 cells was 51,31 and 49 h,respectively.Although these lines were insusceptible to Autographa californica multiple nuclear polyhedrosis virus（AcMNPV） with the infection rate of 33%-40% at 96 h post infection,QB-Ms2-2 cell line showed higher level of secreted SEAP production in comparison with BTI-Tn5B1-4 cell line.In this study,a SEAP high expression type cell line,QB-M2-2 was isolated from the three new cell lines established from M.sexta,which may provide a source for searching novel clones with higher production of recombinant proteins.

关 键 词：烟草天蛾 新细胞系 重组蛋白 生长曲线 苜蓿银纹夜蛾核型多角体病毒 

分 类 号：Q962[生物学—昆虫学]