pGro7 vector (V012802)

Price Information

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

Basic Vector Information

Plasmids of chaperone. The function of the plasmid pGro7 is to express the GroEL and GroES proteins, which are chaperonins involved in protein folding in the cytoplasm of Escherichia coli. pGro7 is a high-copy-number plasmid, meaning that it exists in multiple copies per cell, and it is often used for overexpression of proteins in E. coli. The GroEL and GroES proteins form a complex that helps to fold newly synthesized proteins into their correct conformation, preventing protein aggregation and enhancing protein stability. pGro7 is a valuable tool in molecular biology for protein production and research.

pGro7 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

• Pei C, Peng X, Wu Y, Jiao R, Li T, Jiao S, Zhou L, Li J, Du Y, Qian EW. Characterization and application of active human α2,6-sialyltransferases ST6GalNAc V and ST6GalNAc VI recombined in Escherichia coli. Enzyme Microb Technol. 2024 Mar 12;177:110426. doi: 10.1016/j.enzmictec.2024.110426. Epub ahead of print. PMID: 38503081.

pGro7 vector Sequence

LOCUS       V012802                 5467 bp    DNA     circular SYN 10-JUN-2022
DEFINITION  Exported.
ACCESSION   V012802
VERSION     V012802
KEYWORDS    pGro7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5467)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1862..1915
                     /label="S loop"
                     /note="GroES chaperone mobile loop that interacts with
                     GroEL"
     CDS             2154..3797
                     /gene="groEL"
                     /label="Chaperonin GroEL"
                     /note="Chaperonin GroEL from Escherichia coli O8 (strain
                     IAI1). Accession#: B7M8Q4"
     rep_origin      4076..4620
                     /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        5146..5248
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             join(5249..5467,1..438)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"