Overview:
Defensins are a large family of peptides of which two groups exist in mammals: alpha defensins and beta defensins - which are distinguishable by the spacing and connectivity of the conserved cysteine residues within the mature peptides. It is thought that the function of defensins in the eradication of pathogens from the host system is to insert themselves into the bacterial membrane under the influence of membrane potential, in doing so forms channels which lead to leakage of cytoplasmic molecules and cell death. Unlike hBD-1, this second peptide, human beta-defensin-2 (hBD-2), is regulated at a transcriptional level in response to contact with microorganisms, and is highly effective in killing the gram negative bacteria. Its expression is also upregulated by the cytokine tumor necrosis factor-alpha.
Gene Aliases:
Beta-defensin 4A, BD-2, hBD-2, Defensin, beta 2, Skin-antimicrobial peptide 1, SAP1, DEFB102, DEFB2, DEFB4.
Genbank Number:
O15263
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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。

