Amplification of All Immunoglobulin V Genes of Human Atherosclerosis by Polymerase Chain Reaction / 上海第二医科大学学报

ABSTRACT

Objective To amplify all immunoglobulin V genes of human atherosclerosis by PCR and to sequence the products of PCR cloned into T-vector. Methods Peripheral blood samples of 50 patients with atherosclerosis were collected,from which lymphocytes were segregated by density gradient centrifugation and total RNA was extracted by TRIzol reagent.V genes of VL(including VK and V?) and VH were amplified by RT-PCR,and the products were digested by restriction enzyme and then cloned into T-vector.Two clones were picked randomly to sequence.(Results)Total RNA were extracted purely with integrity,and all V genes of VL and VH were amplified by PCR successfully.The sequences were highly homologous to human immunoglobulin genes. Conclusion All immunoglobulin V genes of human artherosclerosis were amplified successfully,which lays a foundation for the construction and (selection) of phage library.