首页|异色瓢虫海藻糖合成酶基因的克隆及低温诱导表达分析

异色瓢虫海藻糖合成酶基因的克隆及低温诱导表达分析

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海藻糖是昆虫的血糖,在昆虫体内主要通过海藻糖合成酶(trehalose-6-phosphate synthase,TPS)催化合成.本研究通过同源克隆和cDNA末端快速扩增(rapid-amplification of cDNA ends,RACE)技术,从异色瓢虫Harmonia axyridis中克隆得到了TPS基因的cDNA全长序列,命名为HaTPS(GenBank登录号:FJ501960),全长2949 bp,包含3′非翻译区为505 bp,5′非翻译区为26bp,开放阅读框长2418 bp,共编码805个氨基酸.软件分析显示该基因编码蛋白的分子量为90.58 kD,等电点为7.01,包含两个糖基化位点,无信号肽和跨膜结构.同源比对分析发现,昆虫中TPS基因高度保守,包含两个保守的结构域.同时,采用实时荧光定量PCR技术对异色瓢虫HaTPS在不同发育阶段、低温诱导条件下的表达量进行了研究.结果表明:HaTPS在预蛹期的表达量最高；在短时低温诱导条件下,HaTPS的表达量随着温度的降低而显著升高,在降温和升温处理条件下,HaTPS的表达最呈现先升高后下降的表达趋势.结果表明,TPS基因在昆虫抗逆中起到了重要的调节作用.昆虫经过低温诱导,其TPS基因的调控能力得到提升.
Molecular cloning and cold-induced expression of trehalose-6-phosphate synthase gene in Harmonia axyridis (Coleoptera: Coccinellidae)
Trehalose, which is the blood sugar in insects, is synthesized mainly by trehalose-6-phosphate synthase (TPS) in insects. By homologous cloning and rapid-amplification of cDNA ends (RACE), the full cDNA of the TPS gene in Harmonia axyridis was cloned and named HaTPS (GenBanfc accession number; FJ501960). This gene with 5' and 3' non-coding region of 26 bp and 505 bp, respectively, contains an open reading frame of 2 418 nucleotides encoding a protein of 805 ammo acids with the predicted molecular weight of 90.58 kDa and pI of 7.01. The encoded protein contains two potential N-glycosylation sites, without signal peptide and transmembrane domain. The homology comparison showed that insect TPS was highly conserved with two conserved domains. Real-time fluorescent quantitative PCR was conducted at different developmental stages under the cold induction to detect the expression of HaTPS. The results showed the expression level of HaTPS was the highest in the pre-pupal stage. Under the condition of short-term cold induction, the HaTPS expression was dramatically increased with temperature decreasing; and under the condition of warming and cooling, its expression level increased first, and then declined. The results suggest that TPS plays an important role in the regulation of insect resilience and its regulatory capacity is increased significantly under cold-induction.

Harmonia axyridistrehalose-6-phosphate synthase (TPS)trehalosecold inductionrealtime fluorescent quantitative PCR

秦资、王甦、魏苹、徐彩娣、唐斌、张帆

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杭州师范大学生命与环境科学学院,杭州310036

北京市农林科学院植物保护环境保护研究所,北京100097

异色瓢虫 海藻糖合成酶 海藻糖 低温诱导 实时荧光定量PCR

国家重点基础研究发展规划(973计划)国家自然科学基金国家自然科学基金浙江省重点科技创新团队项目杭州市科技局计划项目杭州师范大学优秀中青年教师支持计划浙江省大学生科技创新活动计划杭州师范大学科技创新与文化创意基金杭州师范大学研究生创新基金资助项目

2009CB11920631071731310008802010R5003920091133B08JTAS2011-01-0312010R4210042009XJ007Y20100170

2012

昆虫学报
中国科学院动物研究所,中国昆虫学会

昆虫学报

CSTPCDCSCD北大核心
影响因子：0.756
ISSN：0454-6296
年,卷(期)：2012.55(6)
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