,Polymorphism Analysis and In Vitro Expression of RANTES Genes

• 作 者: ZENG Qingping(曾庆平) YANG Ruiyi(杨瑞仪) FENG Liling(冯丽玲) FU Linchun(符林春)
• 机 构: Tropical Medicine Institute, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405,
• 出 处: 中华性传播感染杂志（英文版）
• 发表日期: 2001年2期
• 关键词: RANTES Gene cloning Polymorphism In vitro expression AIDS

Objective: To clone, sequence and express the primate β-chemokine RANTES genes, hRANTES from H. sapiens and mRANTES from M. Mulatta, in order to explore the possibility of AIDS gene therapy.Methods: hRANTES and mRANTES were amplified by reverse transcription-polymerase chain reaction (RT-PCR)from RNAs extracted from phytoagglutinin (PHA)-activated peripheral blood lymphocytes. hRANTES was cloned,sequenced and expressed in vitro, and mRANTES was directly sequenced for homology comparison.Results: An expected 276 bp fragment was obtained in both amplifications, and sequence data demonstrated a relatively high homology among different copies of hRANTES (97%),and hRANTES was up to 95.6% homologous to mRANTES.When compared with RANTES from other mammals,hRANTES gave rise to a homology ranging from 77% to 86%.The cloned hRANTES was expressed in vitro and a positive signal of RANTES was detected by dot blotting.Conclusion: The full-length of hRANTES sequence was submitted to GenBank and had been released. Our mRANTES sequence is first reported and not yet appeared in GenBank.The successful cloning and expression of hRANTES will provide a basis for AIDS gene therapy in the future.