Zheng Y-P, Krell P J, Doucet D, Arif B M, Feng Q-L
Great Lakes Forestry Center, Sault Ste Marie, Ontario, Canada.
Arch Insect Biochem Physiol. 2008 May;68(1):49-59. doi: 10.1002/arch.20235.
A beta-N-acetylglucosaminidase cDNA (CfGlcNAcase) was cloned from the spruce budworm, Choristoneura fumiferana. Western blotting analysis of developmental CfGlcNAcase expression revealed high levels of expression of the gene on the last day of the 5th instar larvae and the first day in the 6th instar larvae, followed by a decrease to background levels during the intermolt of the 6th instar. CfGlcNAcase was detected again from the last day of the 6th instar to day 2 of pupal stage. CfGlcNAcase expression was induced by tebufenozide at 24 h post treatment and remained at high levels until 72 h. Immunohistochemical localization analysis of CfGlcNAcase indicated that CfGlcNAcase was present in the molting fluid, epidermis, trachea, and hemolymph in prepupae during the transformation from larva to pupa. CfGlcNAcase cDNA was expressed into a recombinant protein in bacterial and baculovirus systems and the protein expressed in the baculovirus system had a higher chitinolytic activity than in the bacterial system and appeared to be secreted.
从云杉芽卷叶蛾（Choristoneura fumiferana）中克隆出了一个β-N-乙酰氨基葡萄糖苷酶cDNA（CfGlcNAcase）。对CfGlcNAcase在发育过程中的表达进行蛋白质免疫印迹分析显示，该基因在5龄幼虫的最后一天和6龄幼虫的第一天有高水平表达，随后在6龄幼虫的蜕皮间期降至背景水平。从6龄幼虫的最后一天到蛹期的第2天再次检测到CfGlcNAcase。在处理后24小时，虫酰肼诱导CfGlcNAcase表达，并且其表达一直维持在高水平直至72小时。CfGlcNAcase的免疫组织化学定位分析表明，在从幼虫向蛹转变的预蛹期，CfGlcNAcase存在于蜕皮液、表皮、气管和血淋巴中。CfGlcNAcase cDNA在细菌和杆状病毒系统中表达为重组蛋白，在杆状病毒系统中表达的蛋白比在细菌系统中具有更高的几丁质分解活性，并且似乎是分泌型的。
