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首页> 外文期刊>Toxicology Letters: An International Journal Providing a Forum for Original and Pertinent Contributions in Toxicology Research >The study on venom proteins of Lapemis hardwickii by cDNA phage display.
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The study on venom proteins of Lapemis hardwickii by cDNA phage display.

机译：用cDNA噬菌体展示技术研究哈氏羊草的毒蛋白。

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In this study, a cDNA T7 phage display library was constructed from sea snake Lapemis hardwickii venom gland mRNA to analyze the components in venom and find new toxins. All the venom gland cDNA-encoding proteins, including housekeeping proteins and venom proteins were expressed on the surface of bacteriophage T7. This library was then panned with rabbit anti-sea snake venom IgG. Phage particles displaying venom-components with interaction with the antibodies were enriched. Thus, phage-carrying venom proteins, such as short chain neurotoxin, long chain neurotoxin, PLA2-like toxin, and c-type lectin-like protein were found, some of them were never reported previously. Four different short chain neurotoxins (SNT-1, 2, 3, 4) were cloned and expressed in Escherichia coli. The LD(50) and analgesic activities of their purified forms were determined. Their structure-function relationship were studied with the aid of a toxin-nAChR complex model constructed by ourselves. Among them, SNT-4 was a new neurotoxin identified in this study and showed potential as pain killer. These results prove that cDNA phage display technique has great advantage than traditional cDNA library method because of the linkage between phenotype and genotype of phage, which provided an effective means to study unknown proteins with target functions.
机译：在这项研究中，从海蛇Lapemis hardwickii毒液腺mRNA构建了cDNA T7噬菌体展示文库，以分析毒液中的成分并发现新的毒素。所有的毒腺cDNA编码蛋白，包括管家蛋白和毒液蛋白，都在噬菌体T7的表面表达。然后用兔抗海蛇毒IgG淘洗该文库。展示具有与抗体相互作用的毒液成分的噬菌体颗粒被富集。因此，发现了携带噬菌体的毒蛋白，例如短链神经毒素，长链神经毒素，类PLA2毒素和c型凝集素样蛋白，其中一些以前从未报道过。克隆了四种不同的短链神经毒素（SNT-1、2、3、4）并在大肠杆菌中表达。确定了其纯化形式的LD（50）和镇痛活性。利用我们自己构建的毒素-nAChR复合物模型研究了它们的结构-功能关系。其中，SNT-4是本研究中鉴定出的一种新的神经毒素，具有潜在的止痛作用。这些结果证明，由于噬菌体的表型和基因型之间的联系，cDNA噬菌体展示技术比传统的cDNA文库方法具有更大的优势，为研究具有靶功能的未知蛋白提供了有效的手段。

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