Sietsma J H, Beth Din A, Ziv V, Sjollema K A, Yarden O
Department of Plant Biology, University of Groningen, Haren, The Netherlands.
Microbiology (Reading). 1996 Jul;142 ( Pt 7):1591-6. doi: 10.1099/13500872-142-7-1591.
Polyclonal anti-chitin synthase antibodies raised against the Saccharomyces cerevisiae CHS2 gene product were used to identify and localize chitin synthase in the filamentous ascomycete Neurospora crassa. A single band of approximately 110 kDa was observed in Western blots of total protein extracts of N. crassa, probed with these antibodies. However, several additional bands were labelled when membrane fraction proteins (microsomes) were probed. Histo-immunochemical localization of chitin synthase confirmed that the polypeptide is compartmentalized in membranous vesicles (chitosomes), which are abundant in the vicinity of the hyphal tip. TEM analysis did not reveal chitin synthase in the plasma membrane. However, dense labelling of membrane-associated chitin synthase was observed by light-microscopic analysis of N. crassa protoplasts and at young hyphal tips.
针对酿酒酵母CHS2基因产物产生的多克隆抗几丁质合酶抗体，被用于在丝状子囊菌粗糙脉孢菌中鉴定和定位几丁质合酶。用这些抗体探测粗糙脉孢菌总蛋白提取物的蛋白质免疫印迹时，观察到一条约110 kDa的单条带。然而，当探测膜部分蛋白（微粒体）时，有几条额外的条带被标记。几丁质合酶的组织免疫化学定位证实，该多肽被分隔在膜泡（几丁质体）中，这些膜泡在菌丝尖端附近大量存在。透射电镜分析未在质膜中发现几丁质合酶。然而，通过对粗糙脉孢菌原生质体和幼嫩菌丝尖端的光学显微镜分析，观察到与膜相关的几丁质合酶有密集标记。
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