Cloning And Function Primary Analysis Of Cold Resistance Relative Gene From The Saussurea Involucrata Kar.et Kir

Saussurea involucrata Kar.et Kir is the special species in xinjiang.It is capable of tolerating severe abiotic stress conditions. Especially it has a high cold resistance.A sikSACPD gene and dehydrin-like gene(sikDh) related with cold stress have been cloned by RT-PCR and bioinformation analysis,construction of full-length cDNA library and their functions have been researched in the transgenic tobacoo.1. Construction of full-length cDNA library from Saussurea involucrata Kar.et Kir by SMART techniqueWe obtained full length cDNA library by SMART technique from cold inducible Saussurea involucrata Kar.et Kir RNA.2. SikSACPD gene was cloned by RT-PCR and PCRa full length cDNA clone SikSACPD(1555bp)by RT-PCR technique from cold inducible Saussurea involucrata Kar.et Kir RNA.3 SikDh , dehydrin-like gene related with cold , drought stress ,was otained by bioinformation analysisClones were selected randomly and sequencing , bioinformation analysis, sikDh , dehydrin-like gene related with cold , drought stress ,was otained. full length cDNA (659bp),CDS:345bp. DsikDh has been cloned from genomic DNA. Sequence analysis shows that there is no intron .4. Expression of sikSACPD,sikDh gene in E.coli BL21Construction of prokaryotic expression vector pTHIOsikSACPD, pTHIOsikDh,and expression of them in E.coli BL21.Recombinant proteins were puritied and inserted into rabbit for preparation of antiserm.