Pcmv blank

Manufactured by Beyotime

Sourced in China

PCMV-Blank is a plasmid vector designed for recombinant protein expression in mammalian cell lines. It contains a cytomegalovirus (CMV) promoter, a multiple cloning site, and a transcription termination sequence. This vector can be used as a backbone for the insertion of target genes of interest for expression in mammalian cells.

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Lab products found in correlation

Pcmv n myctag, by Beyotime (2 mentions) Plvx shrna2 mcherry vector, by New England Biolabs (2 mentions) Plvx shrna1, by Takara Bio (2 mentions) Hemagglutinin ha ubiquitin, by Addgene (2 mentions) Pcmv n flag, by Beyotime (2 mentions) Hifi dna assembly master mix, by New England Biolabs (2 mentions) Q5 site directed mutagenesis kit, by New England Biolabs (2 mentions) Plvx ires zsgreen, by Takara Bio (2 mentions) Pspax2, by Addgene (2 mentions) Plvx shrna2, by Takara Bio (2 mentions) Pcl eco, by Addgene (2 mentions) Pmd2 g, by Addgene (2 mentions) Pmirglo dual luciferase vector, by Promega (1 mentions) Pgl3 basic, by Promega (1 mentions)

5 protocols using pcmv blank

Targeted Regulation of HMGB1 and YY1 via miR-449a

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The miR-449a mimics, YY1-siRNA targeting human YY1, and negative control RNAs for both the miRNA and siRNA were chemically synthesized by GenePharma (GenePharma, Shanghai, China). For the expression vector, HMGB1 and YY1 were cloned into pCMV-Blank (Beyotime, Shanghai, China) between the site BamH1 and site EcoR1. We used a bioinformatics analysis to determine that both HMGB1 and YY1 are targets of miR-449a (TargetScan, www.targetscan.org). The linker fragment containing the HMGB1 or YY1 wild-type or the mutant 3′UTR-binding site was synthesized and cloned into the pmirGLO Dual-Luciferase vector (Promega, Madison, WI, USA) between site Sac1 and site Xho1. All the primer sequence information is given in Additional file 1: Table S1.

Cai Y., Jiang C., Zhu J., Xu K., Ren X., Xu L., Hu P., Wang B., Yuan Q., Guo Y., Sun J., Xu P, & Qiu Y. (2019). miR-449a inhibits cell proliferation, migration, and inflammation by regulating high-mobility group box protein 1 and forms a mutual inhibition loop with Yin Yang 1 in rheumatoid arthritis fibroblast-like synoviocytes. Arthritis Research & Therapy, 21, 134.

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Overexpressing Lsd1 and p62 in Cells

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To overexpress the Lsd1 and p62 gene, Lsd1 and p62 were cloned into pCMV‐Blank (D2602; Beyotime). The promoter of p62 was cloned into pGL3‐Basic (E1751; Promega). Golden star t6 super PCR mix was purchased from Beijing Tsingke Co., Ltd. All of the constructs were verified by sequencing. Primers are listed in Table S1.

He M., Zhang T., Zhu Z., Qin S., Wang H., Zhao L., Zhang X., Hu J., Wen J., Cai H., Xin Q., Guo Q., Lin L., Zhou B., Zhang H., Xia G, & Wang C. (2020). LSD1 contributes to programmed oocyte death by regulating the transcription of autophagy adaptor SQSTM1/p62. Aging Cell, 19(3), e13102.

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Cloning and Verifying Ash1l Gene

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Gene coding area of Ash1l was cloned into pCMV-Blank (D2602; Beyotime, China). Golden star t6 super PCR mix was purchased from Beijing Tsingke Co., Ltd. All of the constructs were verified by sequencing. Primers are listed in Supplemental Table S1.

Zhang T., Ren T., Lin H., Tong Y., Zhang J., Nie J., Zhu Y., Wang Y., Jin B., Zhang C., Chen T, & He M. (2022). ASH1L contributes to oocyte apoptosis by regulating DNA damage. American Journal of Physiology - Cell Physiology, 323(4), C1264-C1273.

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Plasmid Construction and Manipulation

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MSCV-IRES-GFP, MSCV-ICN1-IRES-GFP and MSCV-ShRNA-ICN1-IRES-GFP were kindly provided by Professor Hudan Liu, Medical Research Institute, Wuhan University, Wuhan, China. pLVX-ShRNA1, pLVX-ShRNA2 and pLVX-IRES-ZsGreen vectors were purchased from Clontech Laboratories. Hemagglutinin (HA)-Ubiquitin, pMD2.G, psPAX2 and pCL-Eco were purchased from Addgene. pCMV-Blank, pCMV-N-MycTag and pCMV-N-Flag were purchased from Beyotime Biotechnology. A modified pLVX-ShRNA2-mCherry vector in which the ZsGreen fluorescent marker was replaced by an mCherry fluorescent marker was made using HiFi DNA Assembly Master Mix (NEB, Cat No. E2621L). ShSIRT1 and ShScramble were cloned into pLVX-ShRNA2-mCherry. ShScramble, ShMYC and Shp27-human were cloned into pLVX-ShRNA2. The SIRT1 coding region was cloned into pCMV-N-MycTag, pCMV-N-Flag and pLVX-IRES-ZsGreen. Mutant SIRT1-H363Y vector was generated using Q5® site-directed mutagenesis kit (NEB, Cat No. E0554). Sanger sequencing of the SIRT1-H363Y plasmid verified the presence of the desired mutation (Fig. S3a). CDK2, SKP2, MYC and mCherry were cloned into pCMV-Blank. CDK2 and p27 were cloned into pCMV-N-Flag. Shp27-mouse and ShRen were cloned into MSCV-ShRNA-ICN1-IRES-GFP.

Wang F., Li Z., Zhou J., Wang G., Zhang W., Xu J, & Liang A. (2021). SIRT1 regulates the phosphorylation and degradation of P27 by deacetylating CDK2 to promote T-cell acute lymphoblastic leukemia progression. Journal of Experimental & Clinical Cancer Research : CR, 40, 259.

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Molecular Cloning and Plasmid Construction

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MSCV-IRES-GFP, MSCV-ICN1-IRES-GFP and MSCV-ShRNA-ICN1-IRES-GFP were kindly provided by Professor Hudan Liu, Medical Research Institute, Wuhan University, Wuhan, China. pLVX-ShRNA1, pLVX-ShRNA2 and pLVX-IRES-ZsGreen vectors were purchased from Clontech Laboratories. Hemagglutinin (HA)-Ubiquitin, pMD2.G, psPAX2 and pCL-Eco were purchased from Addgene. pCMV-Blank, pCMV-N-MycTag and pCMV-N-Flag were purchased from Beyotime Biotechnology. A modi ed pLVX-ShRNA2-mCherry vector in which the ZsGreen uorescent marker was replaced by an mCherry uorescent marker was made using HiFi DNA Assembly Master Mix (NEB, Cat No. E2621L). ShSIRT1 and ShScramble were cloned into pLVX-ShRNA2-mCherry. ShScramble, ShMYC and Shp27-human were cloned into pLVX-ShRNA2. The SIRT1 coding region was cloned into pCMV-N-MycTag, pCMV-N-Flag and pLVX-IRES-ZsGreen. Mutant SIRT1-H363Y vector was generated using Q5® site-directed mutagenesis kit (NEB, Cat No. E0554). CDK2, SKP2, MYC and mCherry were cloned into pCMV-Blank. CDK2 and p27 were cloned into pCMV-N-Flag. Shp27-mouse and ShRen were cloned into MSCV-ShRNA-ICN1-IRES-GFP.

Wang F., Li Z., Zhou J., Wang G., Zhang W., Xu J., & Liang A. (2021). SIRT1 Regulates the Phosphorylation and Degradation of P27 by Deacetylating CDK2 to Promote T-cell Acute Lymphoblastic Leukemia Progression.

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