Molecular cloning and expression analysis of the SpbHex(hexamerin gene) in soybean pod borer (Leguminivoraglycinivorella Matsumurav)
Author of the article:ZANG Zhen-Yuan** JIA Bao-Hui LI Han-Zhe LI Na MENG Fan-Li LI Wen-Bin***
Author's Workplace：Key Laboratory of Soybean Biology, Ministry of Education, School of Agriculture, Northeast Agricultural University,Harbin 150030, China
Key Words：soybean pod borer, hexamerin gene, gene clone, expression analysis
Abstract： [Objectives] In insects, Storage Superfamily Proteins (SSPs) accumulate apparently to serve as sources of amino acids during metamorphosis, reproduction and development. Hexamerins are important SSPs. In order to study the function of the hexamerin gene in the development of soybean pod borer, the gene was cloned and its expression analyzed. [Methods] The full length of the cDNA sequence of the soybean pod borer hexamerin gene (SpbHex) was amplified using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The expression of SpbHex in all developmental stages and tissues was analyzed using Realtime quantitative PCR. [Results] A soybean pod borer gene encoding hexamerin (SpbHex) was cloned. The full length of SpbHex was 2 161 bp with an open reading frame (OFR) of 2 112 bp. The ORF of SpbHex encodes a protein of 703 amino acids with a predicted molecular mass of approximately 84.15 ku. RT-PCR analyses indicates that SpbHex was expressed in all development stages, with the highest expression in fourth instar larvae and the lowest in first instar larvae and adults. Meanwhile, RT-PCR analyses indicates high quantities of SpbHex transcript in the fat body and low amounts in the cuticle. [Conclusions] The hexamerin gene of the soybean pod borer was cloned and its transcription profiles determined in all developmental stages and tissues. The present findings lays the foundation for further study of the function of SpbHex in the development of the soybean pod borer.