pGP5-5, pTrx-2 (ATCC® 67287)

Organism: Escherichia coli (Migula) Castellani and Chalmers  /  Clone Type: Clone  /  Depositors: Harvard Medical School, CC Richardson, Harvard Medical School

Permits and Restrictions
Permits and Restrictions

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Designations pGP5-5, pTrx-2
Species Escherichia coli (Migula) Castellani and Chalmers
Depositors Harvard Medical School, CC Richardson, Harvard Medical School
Applications
M13mGP1-2 (ATCC 40303) can coexist in the same cell as the two plasmids of ATCC 67287, pGP5-5 and pTrx-2. The T7 RNA polymerase of M13mGP1-2 can be used to regulate expression of T7 DNA polymerase by inducing the RNA polymerase with IPTG.
pTrx-2 contains two tandem copies of trxA cloned into ptac-12. Expression of the gene is controlled by the tac promoter and is inducible by IPTG. pTrx-2 is ampicillin resistant and contains a unique EcoRI site.
Insert
DNA: genomic
Gene product: DNA polymerase [trxA]
Media 1065 plus ampicillin plus kanamycin
Biosafety Level 1
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--3.2, uncut; HincII--3.4, 2.5, 1.25, 1.1, 0.72(doublet).
For pGP5-5, the insert size is 3.0 kb and the total size of the plasmid is 6.06 kb. For pTrx-2, the insert size is 0.47 kb per copy of the trxA gene and the total size of the plasmid is 3.23 kb.
pGP5-5 contains the replicon P15A. pTrx-2 contains the replicon ColE1.
T7 DNA polymerase consists of two subunits, a protein encoded by T7 gene 5 and Escherichia coli thioredoxin.
M13mGP1-2 (ATCC 40303) can coexist in the same cell as the two plasmids of ATCC 67287, pGP5-5 and pTrx-2. The T7 RNA polymerase of M13mGP1-2 can be used to regulate expression of T7 DNA polymerase by inducing the RNA polymerase with IPTG.
Escherichia coli K38 = HfrC lambda.
pTrx-2 contains two tandem copies of trxA cloned into ptac-12. Expression of the gene is controlled by the tac promoter and is inducible by IPTG. pTrx-2 is ampicillin resistant and contains a unique EcoRI site. It is also available as ATCC 67286.
pGP5-5 contains two segments of bacteriophage T7: nt 5667-6166 (phi1.1A and phi1.1B promoters) and nt 14306-16869 (gene 5). These are cloned into BamHI-HincII of pACYC177. pGP5-5 is kanamycin resistant and contains a unique XhoI site.
Classification Enterobacteriaceae, Escherichia
References

Tabor S, Richardson CC. T7 DNA polymerase. US Patent 4,795,699 dated Jan 3 1989

Charles C Richardson, personal communication

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