Tenebrio molitor抗冻蛋白基因家族cDNA片段的克隆、序列分析及原核表达

doi:10.1360/yc-006-1532

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HEREDITAS ›› 2006, Vol. 28 ›› Issue (12): 1532-1532~1540.doi: 10.1360/yc-006-1532

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Cloning, Sequencing and Prokaryotic Expression of cDNAs for the Antifreeze Protein Family from the Beetle Tenebrio molitor

LIU Zhong-Yuan, WANG Yun, LÜ Guo-Dong, WANG Xian-Lei, ZHANG Fu-Chun, MA Ji

(Key Laboratory of Molecular Biology, College of Life Science and Technology, Xinjiang University Xinjiang Key Laboratory
of Biological Resources and Genetic Engineering, Urumqi 830046, China)

Received:2006-04-25 Revised:2006-07-31 Online:2006-12-10 Published:2006-12-10
Contact: MA Ji

Abstract

Abstract:

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.

CLC Number:

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, 2006, 28(12): 1532-1532~1540.

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Cloning, Sequencing and Prokaryotic Expression of cDNAs for the Antifreeze Protein Family from the Beetle Tenebrio molitor

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