Cloning, Sequencing and Prokaryotic Expression of cDNAs for the Antifreeze Protein Family from the Beetle Tenebrio molitor
Received date: 2006-04-25
Revised date: 2006-07-31
Online published: 2006-12-10
The partial cDNA sequence coding for the antifreeze proteins in the Tenebrio molitor was obtained by RT-PCR. Sequence analysis revealed nine putative cDNAs with a high degree of homology to Tenebrio molitor antifreeze proteins. The recombinant pGEX-4T-1-tmafp-XJ430 was introduced into E. coli BL21 to induce a GST fusion protein by IPTG. SDS-PAGE of the fusion protein demonstrated that the antifreeze protein migrated at a size of 38 kDa. The immunization was performed by intra-muscular injection of pCDNA3- tmafp-XJ430, and then antiserum was detected by ELISA.The titer of the antibody was 1:2 000. Western blotting analysis showed the antiserum was specific against the antifreeze protein. This finding could lead to further investigation of the properties and function of antifreeze proteins.
LIU Zhong-Yuan, WANG Yun, LÜ, Guo-Dong, WANG Xian-Lei, ZHANG Fu-Chun, MA Ji . Cloning, Sequencing and Prokaryotic Expression of cDNAs for the Antifreeze Protein Family from the Beetle Tenebrio molitor[J]. Hereditas(Beijing), 2006 , 28(12) : 1532 -1532~1540 . DOI: 10.1360/yc-006-1532
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