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Molecular Cloning, Sequence Characterization and Expression Analysis of a CD63 Homologue from the Coleopteran Beetle, Tenebrio molitor
Division of Plant Biotechnology, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agriculture and Life Sciences, Chonnam National University, Gwangju 500-757, Korea
National Research Laboratory of Defense Proteins, College of Pharmacy, Pusan National University, Jangjeon Dong, Kumjeong Ku, Busan 609-735, Korea
Department of Life Science and Biotechnology, College of Natural Sciences, Soonchunhyang University, Asan city 336-745, Korea
Division of Applied Entomology, National Academy of Agricultural Science, Rural Development, 61th, Seodun-dong, Gwonseon-gu, Suwon, Gyeonggi-do 441-853, Korea
Department of Biological Science and the Research Institute for Basic Sciences, Hoseo University, Asan 336-795, Korea
* Author to whom correspondence should be addressed.
Received: 6 September 2013; in revised form: 27 September 2013 / Accepted: 3 October 2013 / Published: 15 October 2013
Abstract: CD63, a member of the tetraspanin membrane protein family, plays a pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of a coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic “Cys-Cys-Gly” motif and “Cys188” residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50%–56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcripts are upregulated to the maximum level of 4.5 fold, in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also caused significant increase to the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.
Keywords: molecular cloning; CD63; TM4SF; expression analysis; Tenebrio molitor; immune elicitors
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Patnaik, B.B.; Kang, S.M.; Seo, G.W.; Lee, H.J.; Patnaik, H.H.; Jo, Y.H.; Tindwa, H.; Lee, Y.S.; Lee, B.L.; Kim, N.J.; Bang, I.S.; Han, Y.S. Molecular Cloning, Sequence Characterization and Expression Analysis of a CD63 Homologue from the Coleopteran Beetle, Tenebrio molitor. Int. J. Mol. Sci. 2013, 14, 20744-20767.
Patnaik BB, Kang SM, Seo GW, Lee HJ, Patnaik HH, Jo YH, Tindwa H, Lee YS, Lee BL, Kim NJ, Bang IS, Han YS. Molecular Cloning, Sequence Characterization and Expression Analysis of a CD63 Homologue from the Coleopteran Beetle, Tenebrio molitor. International Journal of Molecular Sciences. 2013; 14(10):20744-20767.
Patnaik, Bharat B.; Kang, Seong M.; Seo, Gi W.; Lee, Hyo J.; Patnaik, Hongray H.; Jo, Yong H.; Tindwa, Hamisi; Lee, Yong S.; Lee, Bok L.; Kim, Nam J.; Bang, In S.; Han, Yeon S. 2013. "Molecular Cloning, Sequence Characterization and Expression Analysis of a CD63 Homologue from the Coleopteran Beetle, Tenebrio molitor." Int. J. Mol. Sci. 14, no. 10: 20744-20767.