ABSTRACT
MP-Rep fusion gene which size is about 1000bp was constructed by recombinant PCR technique.In order to construct fusion gene,Tobacco mosaic virus(TMV) partial movement protein gene(MP) and Cucumber mosaic virus(CMV) partial replicase gene(rep) were ligated.Two copies of MP-Rep fusion gene were ligated with soybean intron in inverted repeat manner,the recombinant fragments were then inserted into binary vector pBIN438 under the control of 35S promoter.Recombinant clone pBIN438-MP-Rep(i/r) which contained two different virus derived genes was constructed.Recombinant clone pBIN438-MP-Rep(i/r) in accord with expected design was certified by restriction endonuclease enzymes digestion and PCR analysis.This approach provides a basis for Broad-spectrum plant virus resistance mediated by RNA Silencing.
ABSTRACT
OBJECTIVE To recombine J chain gene with HNP-1 into a new germicidal(molecule) J-HNP-1,which can connect with pIgR by J chain,so that by using pIgR as a "bridge" the J-HNP-1(germicidal) peptide can be transported into the epithelial cell of mucous membrane to kill the intracellular microorganisms,then the recombinant is inserted into the mammalian expression system.METHODS The J chain and HNP-1 cDNA were amplified from the plasmids respectively by PCR,then the two cDNA fragments were recombined into J-HNP-1 by recombinant PCR.The J-HNP-1 cDNA fragment was inserted into the(mammalian) expression vector pcDNA3.1(-)/Myc-HisC.RESULTS The J-HNP-1 recombinant was obtained by(connection) of J chain and HNP-1 cDNA by PCR.The recombinant J-HNP-1 cDNA was 786bp.CONCLUSIONS The recombinant J-HNP-1 cDNA and the construction of expression vector are the basis for the new bactericidal peptide production.
ABSTRACT
Recombinant PCR applies to fulfill gene recombination by PCR thermal reaction.Over the twenty years,it has branched into three characteristic strategies:splicing by overlapping extension(SOE),jumping polymerase chain reaction(JPCR)and DNA shuffling.Recently,the technique aimed with exploiting natural source of different allele genes is developing up on simplification of experimental procedure,on trap for mutation and variation,and on highthroughput screening with technology of surface display and fluorescent probe.The recombinant PCR is increasesing value in broad range from biological basic research to bioengineering study.
ABSTRACT
The human CD8 molecule consists of two chains,termed ? and ? termed ? and ? chain ?Transfeetion studies have shown the expression of the CD8? gene product gene product is strict-ly dependent on the CD8? gene product and that CD8 is expressed either as an ?/? homodimer or ?/? heterodimer on transfectants.Soluble CD8(sCD8)molecules come from CD8+cells and take an important rule in immunology and some diseases.For the further studies on sCD8 molecule,we have amplified a new gene coding sCD8 ? chain from the cell surface CD8 ? gene,which the membrane—spanning sequence has been spliced off by the method of recombinant PCR.sCD8? gene was cloned into the vector of pBluescript Ⅱ and sequenced and ready for its exprission stud-ies next.