Amplification, Cloning And Expression Of HIV-1 Protease Gene And The Purification And Identification Of Expressed Product

Acquired Immunodeficiency Syndrome (AIDS) presents a major challenge to health authorities in the last quarter of the 20th century and was considered as a "super canner" for there have being reported no patients with AIDS could live beyond five years! In order to be able to take steps to control the epidemic, it is essential to understand the basic biology of the etiologic agent of AIDS, the human immunodeficiency virus (HIV). As HIV reproduce itself in the body, the genome size mRNA of HIV is translated into three polyproteins. Pr^55(?), Pr^?and gp¹⁶⁰. To produce infectious progeny virions from immature virus particles, cleavage of the polyproteins into the smaller mature structural proteins and into the replicaiton enzymes (reverse transcriptase and integrase) are necessary. This cleavage is accomplished by the viral protease. It has been demonstrated that deletion mutants lacking protease sequences can make HIV produing immature and non-infectious virus particles. The mostly nearly reports even have implied HIV Pr play a improtant role in the progressing of AIDS. So, instensive reserching the function, activity and inhibitors of HIV Pr have become the "highway" of HIV molecular biology and the key-step to control AIDS.In order to further study HIV Pr, We first amplification the HIV-1 Pr whole gene from cDNA clone of a HIV—1 strain ARV-2 by polymerase chain reaction (PCR), meanwhile we reconstructured the Pr gene by introducing the EcoRI and HindIII recognition sites into 5 terminal and 3 terminal of HIV-1 Pr gene respe-ctivily. PCR product was firstly cloned into pUCl9 and the cloned Pr gene wasthen inserted into M13rap18 phage for sequence analysis. The result showed that the cloned Pr gene is correct.