Linked Life Data

8142139

Source:http://linkedlifedata.com/resource/pubmed/id/8142139

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
12
pubmed:dateCreated
1994-5-3
pubmed:abstractText
The entire nef gene sequence of HIV-2, NIH-Z strain, has been cloned into the pJL6 expression vector and used for the synthesis of a 23-kDa protein in E. coli. The expressed protein is a fusion between the N-terminal 13 amino acids of the cII gene, 8 amino acids resulting from the ligation procedure, and the 180 amino acids that comprise the HIV-2 Nef sequence from the NIH-Z strain. The bacterially expressed Nef protein has been purified to apparent homogeneity on analytical scale (10-20 micrograms) by a combination of sequential detergent extraction, gel filtration, and reversed-phase high-performance chromatography. The expressed Nef protein is highly susceptible to proteolysis (chymotryptic-like activity) and this property accounts for the low yield obtained by gel filtration and RP-HPLC. Larger amounts (> 100 micrograms) of the purified Nef protein have been produced by a purification procedure that employs sequential detergent extraction, chromatography on Q-Sepharose in the presence of 7 M urea, and chromatography on hydroxylapatite, also in 7 M urea. The purified HIV-2 Nef protein has been used for the production of polyclonal and monoclonal antibodies. The milder method of purification should facilitate structure-function studies of the Nef protein and its role in the life cycle of HIV.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0889-2229
pubmed:author
pubmed:issnType
Print
pubmed:volume
9
pubmed:geneSymbol
nef
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1225-31
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed:year
1993
pubmed:articleTitle
Purification of an Escherichia coli-expressed Nef protein from the human immunodeficiency virus-type 2.
pubmed:affiliation
Nucleic Acid and Protein Synthesis Laboratory, Program Resources, Inc., National Cancer Institute, Frederick Cancer Research and Development Center, Maryland 21701.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.