Molecular characterization of HIV-2 (ROD) protease following PCR cloning from virus infected H9 cells.
A 450 nucleotide sequence corresponding to the nucleotides 1931-2380 of the viral genome (8) was amplified by polymerase chain reaction (PCR) using template DNA prepared from HIV-2 (ROD) infected H9 cells. The sequence codes for HIV-2 protease and its N-terminal flanking peptide. An identical DNA sequence was obtained from three independent PCR amplifications, which differs from the published sequence of HIV-2 (ROD) in 7 nucleotides scattered throughout the region of the cloned DNA. The cloned DNA was expressed in E. coli cells and resulted in the synthesis of a correctly processed HIV-2 protease, which is enzymatically active. Therefore, none of the seven nucleotide changes, which resulted in two amino acid substitutions, affect the autoproteolytic or trans-cleaving activities of the HIV-2 protease.
Cheng
Y S
YS
DuPont Merck Pharmaceutical Company, Wilmington, DE 19880-0328.
Patterson
C E
CE
Rucker
R G
RG
Otto
M J
MJ
Rizzo
C J
CJ
Korant
B D
BD
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000737:chemistry; D004789:Enzyme Activation; D015658:HIV Infections / Q000235:genetics* / Q000382:microbiology; D016333:HIV Protease / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D015498:HIV-2 / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D004279:DNA, Viral; D016333:HIV Protease