Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V012942 | pKJE7 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pKJE7 vector plasmid is a chaperone plasmid. Primarily for facilitating the production and folding of recombinant proteins in bacterial systems, particularly Escherichia coli (E. coli). It has been shown that the co-expression of ScFvs and the chaperone plasmids PG-tf2, ptf16, pGro7, pKJE7, and pG-KJE8 leads to a noticeable increase in the expression of soluble proteins.
- Vector Name:
- pKJE7
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 7192 bp
- Type:
- Packaging assistance
- Replication origin:
- p15A ori
- Host:
- E. coli
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pKJE7 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
- Nazari A, Farajnia S, Zahri S, Bagherlou N, Tanoumand A, Rahbarnia L. Cytoplasmic Chaperones Enhance Soluble Expression of Anti-EGFR huscFv in Escherichia coli. Iran J Biotechnol. 2020 Apr 1;18(2):e2314. doi: 10.30498/IJB.2020.138200.2314. PMID: 33542937; PMCID: PMC7856399.
- Yasamut U, Thongheang K, Weechan A, Sornsuwan K, Juntit OA, Tayapiwatana C. Evaluating the ability of different chaperones in improving soluble expression of a triple-mutated human interferon gamma in Escherichia coli. J Biosci Bioeng. 2024;138(3):232-238. doi:10.1016/j.jbiosc.2024.06.005
- Rani RM, Syngkli S, Nongkhlaw J, Das B. Expression and characterisation of human glycerol kinase: the role of solubilising agents and molecular chaperones [published correction appears in Biosci Rep. 2023 Oct 31;43(10):BSR-2022-2258_COR. doi: 10.1042/BSR-2022-2258_COR]. Biosci Rep. 2023;43(4):BSR20222258. doi:10.1042/BSR20222258
pKJE7 vector Sequence
LOCUS Exported 7192 bp DNA circular SYN 10-OCT-2024 DEFINITION Exported. ACCESSION V012942 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7192) TITLE Direct Submission REFERENCE 2 (bases 1 to 7192) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..7192 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 1787..3700 /gene="dnaK" /label=Chaperone protein DnaK /note="Chaperone protein DnaK from Escherichia coli O1:K1 / APEC. Accession#: A1A766" CDS 3792..4919 /gene="dnaJ" /label=Chaperone protein DnaJ /note="Chaperone protein DnaJ from Escherichia coli (strain K12). Accession#: P08622" CDS 4939..5529 /gene="grpE" /label=Protein GrpE /note="Protein GrpE from Escherichia coli (strain K12). Accession#: P09372" rep_origin 5800..6345 /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin." promoter 6871..6973 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS join(6974..7192,1..438) /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"