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Plasmid is a closed circular double-stranded DNA molecule that is independent of the host chromosome (nucleoid). Possessing the self-replication ability, the plasmid is able to maintain a stable copy in progeny cells, and express the genetic information it carries. In addition, the plasmid vectors typically carry resistance or reporter genes, facilitating selection and tracing. These characteristics confer high utility of plasmids in genetic engineering. The functions of plasmid vectors in the regulation of gene exrepssions include carrying regulatory elements, as well as achieving induced expression and gene silencing etc.

Chromosome DNA and plasmid DNA

Expression of plasmid vectors in eukaryotic cells

Taskcm can provide the following plasmid construction services:

a. Overexpression Plasmid Construction

Gene plasmid map of MAP3K8

b. Site-directed Mutagenesis of Induced Genes (single-base mutations & multi-base mutations)

c. Gene Truncation Plasmids Construction

Truncation of Stat3-full length & Coiled-coiled domain

d. Subclone Plasmid Construction

Subcloning the humanized FOXO1 from the pFlag-CMV4 vector into the DOX-inducible PLVX-Tetone vector

e. Gene Overexpression Mediated by Lentivirus

Triple-plasmid system packed by lentivirus

Fluorescence image of 293T cells infected with MAP3K8 lentivirus for 24 hours

Validation of the successful protein expression by Western Blot

f. Gene Knockdown Achieved by Constructing shRNA Lentiviral Plasmids

Integrating DSC3-shRNA into the pGLV3-H1/GFP shRNA lentiviral vector

C2C12 cells infected with DSC3-shRNA lentivirus for 24 hours

Validation of the effective mRNA knockdown by RT-qPCR Validation of the effective protein knockdown by Western blot

g. Gene Knockout Achieved by CRISPR-Cas9 Technology

Integrating STAT3-sgRNA into lenti-CRISPR-V2 lentiviral vector

Validation of the successful protein knockout by Western blot

Taskcm can provide specified stable cell lines for the overexpression and knockout services.

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