[Construction and expression of a novel bisbicistronic expression vector: pCMV-Myc-IRES-EGFP].
It is often necessary to construct more than one recombinant plasmids when investigating the characteristics, physchemical features and functional mechanisms of genes or proteins. Repeated sub-cloning procedures including design of primers, enzyme digestion, ligation and verification of recombinant plasmids, have to be involved with. For this reason, it has become a tendency to developing new genetic vectors which can be used in multitude of experiments. Therefore, by using pIRES vector as a backbone, here we reported the construction of a mammalian expression vector: pCMV-Myc-IRES-EGFP which contains the N-terminal c-Myc epitope tag and the enhanced green fluorescent protein (EGFP) translated in an IRES-dependent manner. This novel vector can be used to testify the efficiency of cell transfection, to collect successfully transfected cell population via cytometry, to conduct transcription and translation in vitro, to purify target proteins or to trap their interactional proteins. The availability of this vector can facilitate function study of genes.
Yan
Fei
F
College of Life Sciences of Sichuan University, Chengdu 610041, China.
Zhao
Xin-Yu
XY
Deng
Hong-Xin
HX
Wei
Yu-Quan
YQ
D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D019759:Carbon-Nitrogen Lyases; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D016259:Genes, myc / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D051017:Apoptosis Regulatory Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D019759:Carbon-Nitrogen Lyases; C000604991:DESI2 protein, human