利用大肠杆菌DH5α菌株克隆外源融合基因GAPDH-IFN的优化转化

IF 0.4 4区农林科学 Q4 FISHERIES

Indian Journal of Fisheries 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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引用次数: 0

摘要

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

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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来源期刊

Indian Journal of Fisheries FISHERIES-

CiteScore

0.90

自引率

20.00%

发文量

审稿时长

6-12 weeks

期刊介绍： Indian Journal of Fisheries is published quarterly by the Indian Council of Agricultural Research (ICAR), New Delhi. Original contributions in the field of Fish and fisheries science are considered for publication in the Journal. The material submitted must be unpublished and not under consideration for publication elsewhere. Papers based on research which kills or damages any species, regarded as thratened/ endangered by IUCN crieteria or is as such listed in the Red Data Book appropriate to the geographic area concerned, will not be accepted by the Journal, unless the work has clear conservation objectives.