Document 0491
 DOCN  M9610491
 TI    Tools for the production and purification of full-length, N- or
       C-terminal 32P-labeled protein, applied to HIV-1 Gag and Rev.
 DT    9601
 AU    Jensen TH; Jensen A; Kjems J; Department of Molecular Biology,
       University of Aarhus, Denmark.
 SO    Gene. 1995 Sep 11;162(2):235-7. Unique Identifier : AIDSLINE
       MED/96032349
 AB    We have constructed two new vectors for the production of foreign
       proteins in Escherichia coli. The vectors, pGEX-GTH and pET-HTG, produce
       protein fused to glutathione S-transferase (GST) at the N- and
       C-termini, respectively, allowing one-step purification on
       glutathione-Sepharose. Furthermore, they carry the recognition sequence
       (RRASV) for the catalytic subunit of cAMP-dependent heart muscle kinase
       (HMK) at the terminus distal to the GST tag, enabling specific 32P
       labeling in vitro. By positioning the GST and HMK sequences at opposite
       ends of the introduced gene, only full-length fusion protein becomes
       radiolabeled after purification. Avoiding the labeling of shorter fusion
       protein species, often observed in bacterial expression of foreign
       genes, is particularly important for a number of different purposes,
       including protein mobility shift analysis and protein footprinting
       technology.
 DE    Amino Acid Sequence  Base Sequence  Gene Products,
       gag/GENETICS/*ISOLATION & PURIF  Gene Products, rev/CHEMISTRY/*ISOLATION
       & PURIF  *Genetic Vectors  Glutathione Transferases/CHEMISTRY
       HIV-1/*CHEMISTRY  Molecular Sequence Data  Phosphorus Radioisotopes
       Recombinant Proteins/*ISOLATION & PURIF  Support, Non-U.S. Gov't
       JOURNAL ARTICLE

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