Huang Yufen, Krell Peter J, Ladd Tim, Feng Qili, Zheng Sichun
Guangdong Provincial Key Lab of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou, China.
Arch Insect Biochem Physiol. 2009 Jan;70(1):44-56. doi: 10.1002/arch.20272.
Two Choristoneura fumiferana glutathione S-transferase cDNAs (CfGSTs4 and CfGSTd5) were cloned from a cDNA library constructed using mRNA from the midgut cell line, CF-203. These cDNAs encoded two structurally different proteins with a predicted molecular mass of 23 and 24 kDa, respectively. Amino acid sequence analysis indicates that CfGSTs4 and CfGSTd5 contained Sigma and Delta GST domain, respectively. CfGSTs4 cDNA was expressed as a recombinant protein with the same molecular mass as predicted. Semi-quantitative reverse-transcription PCR analyses indicated that both of these genes were expressed in the epidermis, fat body, and midgut of the 6th instar larvae, as well as CF-203 cells. CfGSTs4 was highly and almost constantly expressed in all tissues during the 6th instar stage. There were higher levels of CfGSTs4 protein in the midgut and fat body than in the epidermis. CfGSTd5 was expressed in the fat body when the insects underwent pupal molting and was constantly expressed in the epidermis and midgut during 6th instar development. CfGSTs4 expression was not affected by ecdysone agonist tebufenozide (RH5992), whereas CfGSTd5 expression was slightly suppressed by the compound.
从使用中肠细胞系CF - 203的mRNA构建的cDNA文库中克隆出两种云杉色卷蛾谷胱甘肽S - 转移酶cDNA（CfGSTs4和CfGSTd5）。这些cDNA分别编码两种结构不同的蛋白质，预测分子量分别为23 kDa和24 kDa。氨基酸序列分析表明，CfGSTs4和CfGSTd5分别包含西格玛和德尔塔谷胱甘肽S - 转移酶结构域。CfGSTs4 cDNA表达为一种与预测分子量相同的重组蛋白。半定量逆转录PCR分析表明，这两个基因在6龄幼虫的表皮、脂肪体和中肠以及CF - 203细胞中均有表达。CfGSTs4在6龄期的所有组织中均高度且几乎持续表达。中肠和脂肪体中的CfGSTs4蛋白水平高于表皮。CfGSTd5在昆虫化蛹蜕皮时在脂肪体中表达，在6龄发育期间在表皮和中肠中持续表达。CfGSTs4的表达不受蜕皮激素激动剂虫酰肼（RH5992）的影响，而CfGSTd5的表达受到该化合物的轻微抑制。
