TY - JOUR
T1 - Expression of the moloney murine leukemia virus and human immunodeficiency virus integration proteins inEscherichia coli
AU - Hizi, Amnon
AU - Hughes, Stephen H.
N1 - Funding Information:
We are most grateful to J. Greenhouse for expert technical assistance, to G. Vande Woude, J. Strathern, and D. Garfinkel for helpful discussions, to M. Powers for the preparation of synthetic DNA, and to H. Marusiodis for preparing the manuscript. Research was sponsored by the National Cancer Institute, DHHS, under Contract NO1-CO-74101 with Bionetics Research, Inc.
PY - 1988
Y1 - 1988
N2 - We have constructed expression plasmids containing the genes for the MuLV and the HIV integration proteins. When introduced intoEscherichia coli, these plasmids cause the production of proteins of the expected molecular weight (43K for MuLV, 31 K for HIV). The wild-type MuLV coding region induces the synthesis of large amounts of integration protein; to obtain large amounts of the full-length HIV integration protein inE. coli, it was necessary to modify the coding region to disrupt a sequence that promotes efficient internal translational initiation inE. coli. It is possible to disrupt this sequence without altering the encoded amino acids; the modified plasmid makes large amounts of the full-length protein and small amounts of the internally initiated protein. Both the MuLV and HIV integration proteins require SDS to be solubilized inE. coli extracts; however, following solubilization with SDS and transfer to a nitrocellulose filter, both integration proteins bind DNA.
AB - We have constructed expression plasmids containing the genes for the MuLV and the HIV integration proteins. When introduced intoEscherichia coli, these plasmids cause the production of proteins of the expected molecular weight (43K for MuLV, 31 K for HIV). The wild-type MuLV coding region induces the synthesis of large amounts of integration protein; to obtain large amounts of the full-length HIV integration protein inE. coli, it was necessary to modify the coding region to disrupt a sequence that promotes efficient internal translational initiation inE. coli. It is possible to disrupt this sequence without altering the encoded amino acids; the modified plasmid makes large amounts of the full-length protein and small amounts of the internally initiated protein. Both the MuLV and HIV integration proteins require SDS to be solubilized inE. coli extracts; however, following solubilization with SDS and transfer to a nitrocellulose filter, both integration proteins bind DNA.
UR - http://www.scopus.com/inward/record.url?scp=0024211766&partnerID=8YFLogxK
U2 - 10.1016/0042-6822(88)90128-6
DO - 10.1016/0042-6822(88)90128-6
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AN - SCOPUS:0024211766
SN - 0042-6822
VL - 167
SP - 634
EP - 638
JO - Virology
JF - Virology
IS - 2
ER -
