摘要
乳酸脱氢酶(lactic dehydrogenase,LDH)是糖酵解途径的终止酶,在整个昆虫的发育阶段长期存在。本试验采用同源克隆和锚定PCR技术,从异色瓢虫Harmonia axyridis中克隆到HaxLDH基因的cDNA全序列(GenBank登录号HM362898),全长1336bp,包含214bp的3′非编码区域和123bp的5′非编码区域,可读框长999bp,编码332个氨基酸。通过软件分析,预测该基因编码蛋白的分子量为36.2kD,理论等电点为8.62;包含1个糖基化位点,无信号肽序列和跨膜结构。同源比对不同昆虫的LDH基因cDNA序列,发现HaxLDH拥有8个非常保守的结构域:DLQHG、TAGV/ARQK/RE/DGES/TRL、LVQRN、GSGTNLD、SCHGW/YI/VI/VGEHGD、VWSG/AVNV/IAGV、AYEV/IIK/RLKGYTS/NWAI/VGLS和LSLP。HaxLDH与赤拟谷盗Tribolium castaneum的LDH同源性最高,达67%;系统发育分析也表明二者亲缘关系较近。本研究为探讨LDH在昆虫发育和逆境中的作用奠定了基础。
Lactic dehydrogenase （LDH） of Harmonia axyridis was cloned by homological cloning and anchored PCR. The full length of HaxLDH cDNA sequence is 1336 bp （GenBank accession number HM362898）, which contains a 3＇ untranslated region of 214 bp and a 5＇ untranslated region of 123 bp. HaxLDH cDNA contains an open reading frame of 999 bp, encoding a protein of 332 amino acids residues with calculated molecular mass of 36.20 kD and pI of 8.62. It has one potential N-glycosylation site （residue 352）, with no signal peptide, putative cleavage sites and transmembrane domain. Blast analysis indicated that HaxLDH contains eight conservative domains, DLQHG, TAGV/ARQK/RE/DGES/TRL, LVQRN, GSGTNLD, SCHGW/YI/VINGEHGD, VWSG/AVNV/IAGV, AYEV/IIK/RLKGYTS/NWAI/VGLS and LSLP of LDH family. HaxLDH had 67% identity with that of Tribolium castaneum, and phylogenetic analysis shows that these two species are most closely related.
出处
《中国生物防治学报》
CSCD
北大核心
2011年第4期458-463,共6页
Chinese Journal of Biological Control
基金
国家"973"计划项目(2009CB119206
2006CB102005-3)
国家自然科学基金(31071731
31000880)
杭州市科技局计划项目(20091133B08)
浙江省大学生科技创新活动计划(新苗人才计划)
杭州师范大学生物科学国家特色专业本科生科技创新项目
杭州师范大学优秀中青年教师支持计划(JTAS2011-01-031)
关键词
异色瓢虫
乳酸脱氢酶
基因克隆
序列分析
Harmonia axyridis
lactic dehydrogenase
gene cloning
sequence analysis
