Cloning,sequencing and prokaryotic expression of cDNAs for antifreeze protein family from Beetle Tenebrio molitor  

作　　者：Zhongyuan LIU Yun WANG Guodong LÜ Xianlei WANG Fuchun ZHANG Ji MA 

机构地区：[1]College of Life Science and Technology,Xinjiang University,Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,Urumqi 830046,China

出　　处：《Frontiers in Biology》2008年第3期279-286,共8页生物学前沿（英文版）

摘　　要：Partial cDNA sequences coding for antifreeze proteins in Tenebrio molitor were obtained by RT-PCR.Sequence analysis revealed nine putative cDNAs with a high degree of homology to Tenebrio molitor antifreeze protein genes published in GenBank.The recombinant pGEX-4T-1-tmafp-XJ430 was introduced into E.coli BL21 to induce a GST fusion protein by IPTG.SDS-PAGE analysis for the fusion protein shows a band of 38 kDa.pCDNA3-tmafp-XJ430 was injected into mice to generate antiserum which was later detected by indirect ELISA.The titer of the antibody was 1:2000.Western blotting analysis shows that the antiserum was specifically against the antifreeze protein.Our results laid the foundation for further studies on the properties and functions of insect antifreeze proteins.

关 键 词：Tenebrio molitor antifreeze proteins cDNA fragment sequence analysis prokaryotic expression 

分 类 号：Q78[生物学—分子生物学]