种子特异启动子的克隆和植物表达载体构建.pdfVIP

, Jou rn al ofAnhu iAgr.i Sci. 2010, 38( 1): 63 67 * , , , , , , (暨南大学生物工程学系, 广东广州 510632 ) [], 。8S , , 。[] 8S a 8SGA, 8SGA cDNA 。[] 3 PCR , 472 bp , pBI28SGA2GU S。[], , 。 ; PCR; ; 8S; S188 A 0517 6611( 2010) 01 00063 05 C loning of Seed2specific Promoter and Construction ofPlantTransformation Vector ZHANG M eng2han et al ( D epartment of B ioengin eer ing, Jinan Un iversity, Guangzhou, Guan gdong 5 10632) Abstract [Objective] P romoter is an mi portant tool in p lant gen etic engineerin g, wh ich is critical for develop ing an effic ient p lant b ioreac2 tor. 8S globu lin is the m ost abundant seed storage protein in Vign a radia te, its regu lator sequence may provide a good source of seed2 sp ec ific promoter. [Method] Vigna rad ia te 8S globu lin A subun it promoter sequen ce was amp lified by the method of Genom e2wa lk ing in accordance w ith th e Vigna radiate 8S globu lin A subun it cDNA sequen ce. [Resu lt] Th e amp lified sequ en ce was cloned into pMD182T vector, an d th en tran sformed into E. coli cells. Resu ltant p lasm id was extracted an d subjected to DNA sequ en cing. Th e c lon ed sequence was ana lyzed by var2i ou s promoter prediction servers. Th e sequen ce was shown to contain a numb er of key components of seed2specific promoters; suggesting tha t we h ave successfu lly obtain ed the Vigna rad ia te 8SGA prom oter. [Conclusion ] Sub sequently, 8SGA was sub clon ed into p lant b inary vector pBI121 by rep lac ing CaMV35S promoter up stream of GUS reporter gen e w ith 8SGA promoter, thus prov idin g a basis for investigation of 8SGA promoter function in p lanta. K ey ords P rom oter; PCR; Genome2walk ing; Vigna radia te 8S globu lin; Seed2specific expression vector 植物生物反应器具有诸多优势而被