中华蜜蜂(Apis cerana cerana) MRJP1基因Bacmid表达载体的构建  

The Construction of Bacmid Expression Vector Containing MRJP1 Gene of Apis cerana cerana

作　　者：陶挺[1] 苏松坤[1] 陈盛禄[2] 杜宏沪[1] 

机构地区：[1]浙江大学动物科学学院,浙江杭州310029 [2]浙江大学农业与生物技术学院,浙江杭州310029

出　　处：《蜜蜂杂志》2007年第S1期47-51,共5页Journal of Bee

摘　　要：蜂王浆(RJ,Royal Jelly)是哺育蜂咽下腺与上颚腺分泌的供蜂王和3日龄以内蜜蜂幼虫食用的浆状物质。蛋白质是蜂王浆的重要活性组分,蜂王浆主蛋白MRJPs(Major royal jelly proteins)具有多种生物活性。以中华蜜蜂为材料,通过引物设计和PCR扩增,将中蜂MRJP1的cDNA全长连接到pFastBacHTb转移载体,并将纯化后的pFastBacHTb-MR- JP1质粒转化DH10Bac菌感受态细胞,获得中蜂MRJP1的重组病毒Bacmid表达载体。成功构建的重组病毒Bacmid-MRJP1,可用于转染家蚕细胞,获得重组病毒粒子,得到中蜂MRJP1目的蛋白,为准确测定单一王浆主蛋白MRJP1的生物学功能奠定基础。Royal jelly (RJ) is a thick,milky material that is confirmed to be a secretion product of both hypopharyngeal and mandibular glands of nurse bees.In a colony of bees,the queen is fed on RJ throughout the whole life,while all the other larvae are fed on RJ only for the first 3 days after hatching.Up to now,multiple bioactivities have been found in RJ.The main proteins of RJ are bioactive components and named major royal jelly proteins (MRJPs).MRJP1 is one of the members of MRJPs. With the primers designing and PCR amplifying,the whole cDNA of Apis cerana cerana MRJP1 gene was ligated into the transferring vector pFastBacHTb.Then the plasmid pFastBacHTb-MRJP1 was transformed into competent cells of DHIOBac bacteria and the expression vector of recombinant virus of Bacmid-MRJP1 was constructed successfully.The recombinant virus of Bacmid-MRJPl will be used to transfect cells of silkworm B.mori and produce the target protein MRJPI of Apis cerana cerana in vitro. This would be very helpful to make further researches on the bioactive functions of single member of MRJPs.

关 键 词：中华蜜蜂 蜂王浆 MRJP1 BACMID 表达载体 

分 类 号：S891[农业科学—特种经济动物饲养]