pIRESneo2

质粒类型：哺乳动物表达载体

启动子：CMV

别名：pIRES-neo2，pIRES neo2

载体大小：5295bp

5' 测序引物序列：CMV-F: 5'-CGCAAATGGGCGGTAGGCGTG-3'

3' 测序引物序列：pIRESrv: 5'-GCCCTAGATGCATGCTCG-3'

载体抗性：Ampicillin (氨苄青霉素)

筛选标记：Neomycin (新霉素)

pIRESneo2 contains the internal ribosome entry site (IRES) of the encephalomyocarditis virus (ECMV), which permits the translation of two open reading frames from one messenger RNA (1–3). After selection with G418, nearly all surviving colonies will stably express the gene of interest, thus decreasing the need to screen large numbers of colonies to find functional clones. To select for cells that express high levels of the gene of interest, the selective pressure for antibiotic resistance was increased by moving the initiation codon of the neomycin phosphotransferase gene to a less optimal position for translation as directed by the IRES sequence (attenuated IRES; 1). The expression cassette of pIRESneo2 contains the human cytomegalovirus (CMV) major immediate early promoter/enhancer followed by a multiple cloning site (MCS) that precedes stop codons in all three reading frames, a synthetic intron known to enhance the stability of the mRNA (4), the ECMV IRES followed by the neomycin phosphotransferase (NPT II) gene, and the polyadenylation signal of the bovine growth hormone. Ribosomes can enter the bicistronic mRNA at the 5' end to translate the gene of interest and at the ECMV IRES to translate the antibiotic resistance marker.