Overview:
TNF-alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily and it is secreted predominantly by monocytes/macrophages that have effects on lipid metabolism, coagulation, insulin resistance, and endothelial function (1).TNF-alpha is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation and implicated in a variety of diseases. TNF-alpha also participates in the homeostatic activity-dependent regulation of synaptic connectivity and play roles in synaptic plasticity and modulating responses to neural injury (2).
Gene Aliases:
TNF, TNFa, DIF, TNFA, TNFSF2
Genbank Number:
P01375
References:
1. Shirai, T.et.al: Cloning and expression in Escherichia coli of the gene for human tumour necrosis factor. Nature 313: 803-806, 1985.2. Stellwagen D. et al: Synaptic scaling mediated by glial TNF-alpha. Nature. 2006 Apr 20; 440(7087):1054-9.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
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。

