
Overview:
Vimentin is a member of intermediate filament family of proteins and is an important structural feature of eukaryotic cells (1). Along with microtubules and actin microfilaments, Vimentin make up the cytoskeleton component of cells. Studies have shown that Vimentin is attached to the nucleus, endoplasmic reticulum and mitochondria, either laterally or terminally. Vimentin plays a significant role in supporting and anchoring the position of the organelles in the cytosol (2). Thus, Vimentin plays a key role in maintaining cell shape, integrity of the cytoplasm, and stabilizing cytoskeletal interactions
Gene Aliases:
VIM, FLJ36605
Genbank Number:
NM_003380
References:
1. Fuchs, E., et al: Intermediate filaments: structure, dynamics, function, and disease". Annu Rev Biochem,1994; 63: pp. 345-82. 2. Katsumoto, T. et al: The role of the vimentin intermediate filaments in rat 3Y1 cells elucidated by immunoelectron microscopy and computer-graphic reconstruction. Biol Cell,1990; 68 (2): pp. 139-46.
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实验非常不顺,想构建CDNA文库,但是从mRNA开始屡次失败,考虑主要是纯化过程中损失过多。想从总RNA入手,但是不知道实验步骤。不知那位大侠能提供总RNA建库的实验步骤。另外我现在手头有OligoDT,RT酶,苦于没有第二连合成试剂盒,不知道能否用普通PCR试剂盒Teq酶替代第二链合成过程中的DNA聚合酶I。好像有种方法合成第二链时,不需要另外的引物,利用降解的RNA作引物即可,我想直接设定PCR两个循环,合成第二链,不知方法可行否?愁啊,等着毕业,时间紧急,恳请帮忙。谢谢。

