Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore 54590, Pakistan.
The University of Lahore, Lahore 54590, Pakistan.
Biomed Res Int. 2017;2017:5046451. doi: 10.1155/2017/5046451. Epub 2017 Aug 10.
A thaumatin-like protein gene from Basrai banana was cloned and expressed in . Amplified gene product was cloned into pTZ57R/T vector and subcloned into expression vector pET22b(+) and resulting pET22b-basrai TLP construct was introduced into BL21. Maximum protein expression was obtained at 0.7 mM IPTG concentration after 6 hours at 37°C. Western blot analysis showed the presence of approximately 20 kDa protein in induced cells. Basrai antifungal TLP was tried as pharmacological agent against fungal disease. Independently Basrai antifungal protein and amphotericin B exhibited their antifungal activity against ; however combined effect of both agents maximized activity against the pathogen. Docking studies were performed to evaluate the antimicrobial potential of TLP against by probing binding pattern of antifungal protein with plasma membrane ergosterol of targeted fungal strain. Ice crystallization primarily damages frozen food items; however addition of antifreeze proteins limits the growth of ice crystal in frozen foods. The potential of Basrai TLP protein, as an antifreezing agent, in controlling the ice crystal formation in frozen yogurt was also studied. The scope of this study ranges from cost effective production of pharmaceutics to antifreezing and food preserving agent as well as other real life applications.
从 Basrai 香蕉中克隆并表达了一种娄胎素样蛋白基因。扩增的基因产物被克隆到 pTZ57R/T 载体中,并亚克隆到表达载体 pET22b(+)中,得到的 pET22b-basrai TLP 构建体被引入 BL21。在 37°C 下 6 小时后,在 0.7mM IPTG 浓度下获得最大蛋白表达。Western blot 分析显示,诱导细胞中存在约 20kDa 的蛋白。尝试将 Basrai 抗真菌 TLP 作为药理学制剂用于抗真菌病。独立的 Basrai 抗真菌蛋白和两性霉素 B 对 表现出抗真菌活性;然而,两种药物的联合作用最大限度地提高了对病原体的活性。通过探测靶向真菌菌株质膜麦角固醇与抗真菌蛋白的结合模式,进行对接研究以评估 TLP 对 的抗菌潜力。冰晶形成主要损害冷冻食品;然而,添加抗冻蛋白会限制冷冻食品中冰晶的生长。还研究了 Basrai TLP 蛋白作为冷冻抑制剂在控制冷冻酸奶中冰晶形成的潜力。本研究的范围从具有成本效益的药物生产到抗冻和食品保鲜剂以及其他实际应用。