Optimisation of transformation in the heterologous fusion gene GAPDH-IFN cloning using DH5α strain of Escherichia coli

Pub Date : 2023-06-30 DOI:10.21077/ijf.2023.70.2.123116-19
Anisha Valsalam, K. V. Rajendran, Pooja Vinde, Megha Kadam Bedekar
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Abstract

DNA vaccines are the most essential tool of the disease prevention strategy. In the present study, cloning of a heterologous fusion gene involving glyceraldehyde 3-phosphate dehydrogenase and interferon gamma (GAPDH-IFN) was conceptualised. Cloning was tried using four distinct transformation techniques viz. InstAclone PCR Cloning kit (Fermentas, USA); CaCl2 transformation protocol; Clontech stellar competent cells protocol and PEG 8000-mediated transformation method, for the heterologous GAPDH-IFN fusion gene using DH5α strain of Escherichia coli. The first three methods were found to be unsuitable, and the PEG 8000-mediated transformation method yielded positive clones. Keywords: DNA vcaccine, Edwardsiella tarda, Fusion genes, Labeo rohita, PEG 8000, Transformation methods
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利用大肠杆菌DH5α菌株克隆外源融合基因GAPDH-IFN的优化转化
DNA疫苗是疾病预防战略中最重要的工具。在本研究中,克隆涉及甘油醛3-磷酸脱氢酶和干扰素γ (GAPDH-IFN)的异源融合基因是概念化的。使用四种不同的转化技术进行克隆:instclone PCR克隆试剂盒(Fermentas, USA);CaCl2转换协议;Clontech星态细胞方案和PEG 8000介导转化方法,利用大肠杆菌DH5α菌株进行外源GAPDH-IFN融合基因的转化。前三种方法均不适合,peg8000介导的转化方法获得阳性克隆。关键词:DNA疫苗,迟缓爱德华氏菌,融合基因,罗希塔Labeo, peg8000,转化方法
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