Molecular Cloning and Expression Analysis of Hemolin Gene in Tea Geometrid (Ectropis obliqua)

YU Yugeng; YUAN Zhijun; YIN Kunshan; FU Jianyu; XIAO Qiang

doi:10.13305/j.cnki.jts.2015.04.001

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Journal of Tea Science ›› 2015, Vol. 35 ›› Issue (4) : 307-315. DOI: 10.13305/j.cnki.jts.2015.04.001

Molecular Cloning and Expression Analysis of Hemolin Gene in Tea Geometrid (Ectropis obliqua)

YU Yugeng^1,2, YUAN Zhijun¹, YIN Kunshan¹, FU Jianyu^1,2, XIAO Qiang^1,*

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Abstract

Hemolin, a unique immune protein, belongs to the immunoglobulin superfamily of Lepidoptera insects. A full length cDNA sequence of hemolin gene was cloned by RACE-PCR from tea geometrid (Ectropis obliqua Prout), which was named as EoHML (GenBank Accession No. KM885983). The results of bioinformatics characterization showed that the full length cDNA of EoHML was 1β772βbp, which contained an intact CDS of 1β239βbp and encoded 412 amino acid residues with a putative molecular mass of 45.8βkD and an isoionic point of 8.297. The deduced amino acid sequence demonstrates it belongs to a typical secretory protein that contains 4 Ig functional conservative areas and 2 N-glycosylation sites. Phylogenetic analysis showed that EoHML share the far genetic distance with published hemolin gene and the highest similarity with that of Manduca sexta was only 53%. The results of quantitative real-time PCR analysis further revealed a significant up-regulated expression about 36.5 times in the larvae infected E.obliqua Nucleopolyhedrovirus (EoNPV) than the controls. The study indicated that the EoHML gene may participate in the immune response to EoNPV in tea geometrid.

Key words

expression analysis / gene cloning / hemolin / tea geometrid (Ectropis obliqua)

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YU Yugeng, YUAN Zhijun, YIN Kunshan, FU Jianyu, XIAO Qiang. Molecular Cloning and Expression Analysis of Hemolin Gene in Tea Geometrid (Ectropis obliqua)[J]. Journal of Tea Science. 2015, 35(4): 307-315 https://doi.org/10.13305/j.cnki.jts.2015.04.001

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