Establishment of optimum DDRT-PCR system and screening of antennae differentially expressed genes of Anoplophora glabripennis (Coleoptera:Cerambycidae) | 光肩星天牛mRNA差异显示体系的建立与触角差异表达基因的筛选

Chinese. 建立一套适合光肩星天牛的mRNA差异显示(DDRT-PCR)体系，并对光肩星天牛触角差异表达基因进行初步研究。以光肩星天牛雌、雄个体的触角和去触角的身体部分为材料，对影响mRNA差异显示体系的主要因子进行优化，并研究了其差异表达基因。建立了适合光肩星天牛mRNA差异显示的反应体系，并筛选得到最适的PCR退火温度。使用此体系分离得到28条触角差异表达基因片段，其中9个片段的翻译产物分别与觅食相关蛋白、MYB转录因子、β-乙酰葡萄糖胺转移酶、化学感受蛋白11、表皮蛋白peritrophins 1-I、甘油激酶、抗菌肽Alo-3、26S蛋白酶体(非ATP酶亚基12)和清道夫受体等已知蛋白有较高的同源性。利用建立的DDRT-PCR体系对供试材料进行扩增，扩增产物条带丰富且清晰，可用于分离差异表达基因，得到的触角差异显示基因可能参与了光肩星天牛的嗅觉识别过程。

English. The research was done in order to establish a method of mRNA differential display PCR which was suitable to study the tissue of Anoplophora glabripennis and its antennae differentially expressed genes were also studied. Taking antennae and deantennated bodys of male and female A. glabripennis as material, the critical factors of DDRT-PCR were optimized and differentially expressed genes were analyzed. A special mRNA differential display PCR system for A. glabripennis was established and the suitable annealing temperatures were also found. Using this system, 28 antennae differentially expressed genes’ fragments were obtained and 9 of them showed high sequence similarity to known gene products, including foraging-like protein, MYB transcription factor, beta-1,3-GlcNAc transferase (bre5), chemosensory protein 11, cuticular protein analogous to peritrophins 1-I, glycerol kinase, antimicrobial peptide Alo-3, proteasome 26S non-ATPase subunit 12 and scavenger receptor class B. The production of amplification shows the characteristics of well specificity and distinct DNA fragments under the system which can be used to identify differentially expressed genes in A. glabripennis and the obtained genes may be involved in olfactory recognition.