Molecular cloning of actinohivin, a novel anti-HIV protein from an actinomycete, and its expression in Escherichia coli

Syncytium-inducing variants of the HIV-I virus are correlated with poor dia gnosis and rapid disease progression. We have recently discovered a novel a nti-HIV protein, referred to as actinohivin, that inhibits syncytium format ion. Here we describe the cloning and sequencing of the gene encoding actin ohivin from the actinomycete strain K97-0003, and its expression in Escheri chia coli. The actinohivin gene was located on a 0.8-kb BamHI fragment of g enomic DNA. The fragment contained an open reading frame of 480 bp, which e ncoded a protein of 160 amino acids with calculated molecular weight of 174 92.7. The N-terminal region was found to be a typical signal peptide of pro karyotes, and actinohivin was located at amino acid positions 46-160. The a ctinohivin gene could be expressed in E. coli using a pET30Xa/LIC expressio n vector and the purified recombinant actinohivin was found to inhibit sync ytium formation to a similar extent as actinohivin from its natural source. (C) 2001 Academic Press.