pLP/VSVG vector (V010445)

Price Information

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

This is a lentiviral packaging plasmid for expression of the vesicular stomatitis virus G glycoprotein.

pLP/VSVG vector Map

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

References

• Sun L, Zhang Y, Huang CX, Qu XL, Zhang Y, Zhang JC, Wei X, Zhuang Y, Zhai S, Peng MJ, Jiang H, Hao CQ, Li YH, Lu J, Lian JQ, Sun YT, Bai XF. Therapeutic effect of RANTES-KDEL on inhibition of HIV-1 in CD34(+) human hematopoietic stem cells (hHSC). J Virol Methods. 2008 Dec;154(1-2):194-9.

pLP/VSVG vector Sequence

LOCUS       Exported                5823 bp DNA     circular SYN 02-SEP-2024
DEFINITION  Lentiviral packaging plasmid for expression of the vesicular 
            stomatitis virus G glycoprotein.
ACCESSION   .
VERSION     .
KEYWORDS    pLP VSVG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5823)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5823)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5823)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..458
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        459..662
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          796..1271
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     CDS             1346..2878
                     /label=VSV-G
                     /note="vesicular stomatitis virus G glycoprotein"
     polyA_signal    3083..3477
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     rep_origin      complement(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4749..5606)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5607..5711)
                     /label=AmpR promoter