Cloning of rainbow trout (Oncorhynchus mykiss) histone H3 promoter and the activity analysis in rare minnow (Gobiocypris rarus )  

作　　者：MAOWeifeng SUNYonghua WANGYaping WUGang CHENShangping ZHUZuoyan 

机构地区：[1]StateKeyLaboratoryofFreshwaterEcologyandBiotechnology,InstituteofHydrobiology,ChineseAcademyofSciences,Wuhan430072,China

出　　处：《Progress in Natural Science:Materials International》2004年第4期322-326,共5页自然科学进展·国际材料（英文版）

基　　金：Supported by the Major State Basic Research Development Program of China (Grant No. G2000013109)

摘　　要：Rainbow trout histone H3 (RH3) promoter was cloned via high fidelity PCR. The cloned RH3 promoter was inserted into a promoter-lacked vector pEGFP-1, resulting in an expression vector pRH3EGFP-1. The linearized pRH3EGFP-1 was microinjected into fertilized eggs of rare minnows and the sequential embryogenetic processes were monitored under a fluorescent microscope. Strong green fluorescence was ubiquitously observed at as early as the gastrula stage and then in various tissues at the fry stage. The results indicate that RH3 promoter, as a piscine promoter, could serve in producing transgenic Cyprinoid such as rare minnow. Promoter activity of RH3, CMV and common carp β-actin (CA) were compared in rare minnow by the expression of respective recombinant EGFP vectors. The expression of pCMVEGFP occurred earlier than the following one, pRH3EGFP-1, and then pCAEGFP during the embryogenesis of the transgenics. Their expression activities demonstrated that the CMV promoter is the strongest one, followed by the CA and then the RH3.Rainbow trout histone H3 (RH3) promoter was cloned via high fidelity PCR. The cloned RH3 promoter was inserted into a promoter-lacked vector pEGFP-1, resulting in an expression vector pRH3EGFP-1. The linearized pRH3EGFP-1 was microinjected into fertiilzed eggs of rare minnows and the sequential embryogenetic processes were monitored under a fluorescent microscope. Strong green fluorescence was ubiquitously observed at as early as the gastrula stage and then in various tissues at the fry stage. The results indicate that RH3 promoter, as a piscine promoter, could serve in producing transgenic Cyprinoid such as rare minnow. Promoter activity ofRH3, CMV and common carp ^-actin (CA) were compared in rare minnow by the expression of respective recombinant EGFP vectors.The expression of pCMVEGFP occurred earlier than the following one, pRH3EGFP-1, and then pCAEGFP during the embryogenesis of the transgenics. Their expression activities demonstrated that the CMV promoter is the strongest one, followed by the CA and then the RH3.

关 键 词：enhanced green fluorescent protein (EGFP) histone H3 promoter transgenic fish rare minnow. 

分 类 号：Q959.468[生物学—动物学] Q785