Overview:
PDE9A is a member of the phosphodiesterase (PDE) family that catalyzes the hydrolysis of cGMP to the monophosphate and thus plays an important role in signal transduction by regulating the intracellular concentration of cyclic nucleotides (1). Multiple transcript variants encoding several different isoforms have been found for PDE9A (2). These mRNAs differ in the use of specific combinations of exons located at the 5' end of the gene while the 3' half, encoding the catalytic domain of the protein, always has the same combination of exons. PDE9A has been used in the study of gene- disease association, gene-environment interaction, and pharmacogenomic/toxicogenomic. PDE9A is mainly expressed in spleen, small intestine, and brain.
Gene Aliases:
FLJ90181; HSPDE9A2
Genbank Number:
NM_001001567
References:
1. Fisher, D A. et al: Isolation and characterization of PDE9A, a novel human cGMP-specific phosphodiesterase. J. Biol. Chem. 273: 15559-15564, 1998.2. Soderling, S H. et al: Identification and characterization of a novel family of cyclic nucleotide phosphodiesterases. J. Biol. Chem. 273: 15553-15558, 1998.
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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。

