Overview:
AMPK is a heterotrimer protein kinase consisting of a α catalytic subunit, and non-catalytic β and γ subunits. AMPK is an important energy-sensing enzyme that monitors cellular energy status (1). In response to cellular metabolic stresses, AMPK is activated and phosphorylates and inactivates acetyl-CoA carboxylase (ACC) and beta-hydroxy beta-methylglutaryl-CoA reductase (HMGCR), key enzymes involved in regulating biosynthesis of fatty acid and cholesterol (2).
Gene Aliases:
Subunit A1: PRKAA1, MGC33776, MGC57364Subunit B1: PRKAB1, AMPK, HAMPKb, MGC17785Subunit G1: PRKAG1, AMPKG, MGC8666
Genbank Number:
NM_006251NM_006253NM_002733
References:
1. Minokoshi, Y. et al:AMP-kinase regulates food intake by responding to hormonal and nutrient signals in the hypothalamus. Nature 428: 569-574, 2004.2. Hardie, D G. et al: The AMP-activated protein kinase--fuel gauge of the mammalian cells Eur J Biochem. 1997 Jun 1;246(2):259-73.
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多谢……
各位大侠:小弟最近做个长片段的CDNA与T载体的连接,片段大小6.4K,载体选用的是pUC19和pGEMT-easy,反转录酶用的是RevertAidTirstStrandcDNASynthesisKit(#K1621),CDNA第二链的扩增用的是LATaq。我是用纯病毒的RNA做模板,操作步骤严格按照说明做的,也曾经有一次获得过电泳图清晰显示的是全长6.4KB左右的片段,与载体连接,结果筛选到的几个克隆,插入的片段仅有2KB左右,奈何?可最近几次再用此反转录试剂盒却不能得到所需片段。在此,小弟想问问大家,你们是否遇到同样的问题,是如何解决的?所用的试剂(载体和酶的选择)和方法是什么,可以告诉后来者,共同进步吧。

