The Effect On IFN-γ Gene Expression Of Xinjiang Duolang Sheep By Gossypol And Constuction Of Eukaryotic Expression Vector PcDNA3.1-INF-γ And Transfection Of Lymphocytes

Gossypol,also known as cotton toxins or cottonseed cake,is a polyphenol compound formedin the cotton plants in various parts of plant Brown punctate Pigment Glands,and cottonseed cakeis a kind of high yield,low price,rich in protein animal feed,because of toxicity limited the thelarge-scale application.It has toxicity to ruminant animals or non ruminant animals and showsdose dependent on the animal’s brain,heart,liver,blood vessels and other organ,causing livestockabortion.IFN-γ is a cytokine secreted by immune cells,is an important immuneregulatory factor.Inorder to investigate the effect of gossypol on the immune system and the role of IFN-γ in thegossypol action on the immune system.in this experiment.we used Xinjiang Duolang sheep as theexperimental object,isolated peripheral blood lymphocytes from sheep,co-cultured lymphocyteswith different concentrations of gossypol in vitro,used the method of fluorescence quantitativePCR to detect the ralative expression of IFN-γ.The result showed that With the increasing ofgossypol concentrations,the expression of IFN-γ also showed growing trend.Amplificated the CDS region of IFN-γ by using the method of PCR,used HindIII and BamHIto digest IFN-γ and pcDNA3.1,connected,transformed,sequenced and sequence analysised,theresult showed that successfully cloned a IFN-γ CDS region,the length was455bp,the consistencyis up to more than99%by blastp.Further separated the peripheral blood lymphocytes and culturedlymphocyte in vitro.used electric transfection system to transfect pcDNA3.1-IFN-γ eukaryoticexpression vector into lymphocytes,quantified the expression by using the method of semiquantitative method.The result showed that by comparing the transfection group and the controlgroup of IFN-γ expression,transfection group is significantly higher than that of control group.