
Overview:
CHIP(STUB1) or STIP1 homology and U-box containing protein 1 (also known as E3 ubiquitin protein ligase) is a ubiquitin ligase/cochaperone that participates in protein quality control by targeting a broad range of chaperone protein substrates for degradation (1). CHIP acts as a molecular switch for the degradation of misfolded proteins via the proteasome and lysosome pathways (2). CHIP controls the cellular level of BER enzymes and, correspondingly, the efficiency and capacity of BER. CHIP E3 ubiquitin ligases act sequentially in ER membrane and cytosol to monitor the folding status of CFTR and delF508.CHIP possesses intrinsic E3 ubiquitin ligase activity and promotes ubiquitylation.
Gene Aliases:
CHIP; UBOX1; HSPABP2; NY-CO-7; SDCCAG7
Genbank Number:
NM_005861
References:
1. Min, J.-N. et.al: CHIP deficiency decreases longevity, with accelerated aging phenotypes accompanied by altered protein quality control. Molec. Cell. Biol. 28: 4018-4025, 2008.2. Shin, Y.et.al: The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways. J. Biol. Chem. 280: 23727-23734, 2005.
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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。

