Document Detail


Cloning of cDNA encoding rat spermatidal protein TP2 and expression in Escherichia coli.
MedLine Citation:
PMID:  8954887     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Rat spermatidal protein TP2 is a basic nuclear protein containing two atoms of zinc bound per molecule. We report here cloning of complementary DNA encoding rat TP2 by the RT-PCR method. The nucleotide sequence of cloned TP2 cDNA differs at a few positions from the sequence already reported in the literature. We have cloned rat TP2 cDNA into the expression vector pTrc 99A. Upon induction with 1 mM IPTG, there was a low level of expression of TP2 which could be recovered in the soluble form. Recombinant TP2 was purified from the soluble form. Recombinant TP2 was purified from the soluble extract of E. coli using nickel-agarose and heparin-agarose chromatography and was shown to be identical to native rat TP2 as revealed by immunoblotting with anti-rat TP2 antibodies and radioactive 65Zn-blotting.
Authors:
A R Meetei; M R Rao
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Protein expression and purification     Volume:  8     ISSN:  1046-5928     ISO Abbreviation:  Protein Expr. Purif.     Publication Date:  1996 Dec 
Date Detail:
Created Date:  1997-03-18     Completed Date:  1997-03-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9101496     Medline TA:  Protein Expr Purif     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  409-15     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Indian Institute of Science, Bangalore, India.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/U52958
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Base Sequence
Blotting, Western
Chromosomal Proteins, Non-Histone / genetics*
Cloning, Molecular
DNA, Complementary / chemistry
Electrophoresis, Polyacrylamide Gel
Escherichia coli
Humans
Male
Molecular Sequence Data
Polymerase Chain Reaction
Rats
Zinc
Chemical
Reg. No./Substance:
0/Chromosomal Proteins, Non-Histone; 0/DNA, Complementary; 0/spermatid transition proteins; 7440-66-6/Zinc

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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