The Rapid Screening For Chinese Herbal Medicine Against HIV

AIM: AIDS or "Acquired Immunodefiency Syndrome" is caused by the human immunodeficiencyvirus (HIV), which takes the central cell of the body's immune system-T4lymphocytes as targets, weakensand then destroys the body's immune system, thereby enables the body to become the carrier of variousdiseases, easily contracting all kinds of complications or tumor and to death. AIDS with high mortality ratespreading all over the world is a major threat to development and to human society. Nowadays, thecommonly used drugs in the treatment of AIDS is the protease inhibitor and the reverse transcriptaseinhibitor. These drugs have some defects such as high costs, side effects, easy to cause a variety ofcomplications, and can not take into account of both the fighting against HIV and the patient's immunesystem. Long term treatment always makes patients to develop resistance to these durgs, because the geneof HIV-1enhances mutation after attacked by drugs, and the body produces corresponding antibodies.Themain reason leading to the death of AIDS patients is that the immune system is badly damaged, and patientssuffer from various opportunistic infections and to death.However, the traditional Chinese medical sciencecures from the holistic situation and aims to improve the body's immunity.And traditional Chinese medicineside effects do not cause a variety of complications, at the same time because of the wide range oftraditional medicinal herbs, inexpensive, thus avoiding expensive Western medicine treatment, toxicity,resistance shortcomings, its prospect.The prospect for the development of the traditional Chinese medicineis reasonably cheerful because of the little side effects, effecting no complications, wide-ranging source andlow cost.With the discovery of a new target for the treatment to AIDS---P-TEFb, once we get thedrugs,which can combine with CyclinT1, prevent the combination of CyclinT1and Tat,thus inhibit theactivity of P-TEFb, and thereby block the replication of the HIV genome, we can ensure that the drugs iseffective for replying to the long-term attacking of the HIV-1.At that time, the problem of pesticideresistance will be solved.In this thesis, we constructed the expression vector in eukaryote, used the model organism--Saccharomyces cerevisiae, utilized the yeast two-hybrid technology, dependented on the newtarget-P-TEFb of the treatment to AIDS and screened out some kinds of Chinese herbal medicine preventing the combination of CyclinT1and Tat and inhibiting the activity of P-TEFb.All of that lay a solidfoundation for the medicine used in the clinical treatment.METHODS: Connecte the target gene Cyclint1with the vector pGBKT7and Tat withpGADT7-ReC,and then construct the expression vector in eukaryote--pGBKT7-CyclinT1andpGADT7-ReC-Tat;Transforme the vectors into Saccharomyces cerevisiae,use the yeast two-hybridtechnology to screen out the Chinese herbal medicine;CONCLUSIONS:1In this study,we amplified the correct target gene Cyclint1and Tat by PCR,and successfullyconstructed the eukaryotic expression vector pGBKT7-CyclinT1and pGADT7-ReC-Tat utilizing molecularbiological methods.2We transferred the eukaryotic expression vector pGBKT7-CyclinT1and pGADT7-ReC-Tat intothe Saccharomyces cerevisiae; It was proved that there is interaction betwween them, and then wesuccessfully constructed the yeast two-hybrid system.3Using yeast two-hybrid technology, We successful screened out several Chinese medicines fromthe20kinds of test samples, which could prevent the combination betwween CyclinT1and Tat. They'repokeberry root, Scutellaria baicalensis, Phellodendron and Placenta Hominis.