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首页> 外文期刊>African Journal of Biotechnology >Isolation, identification and PCR amplification of merA gene from highly mercury polluted Yamuna river
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Isolation, identification and PCR amplification of merA gene from highly mercury polluted Yamuna river

机译：汞污染严重的亚木那河中merA基因的分离，鉴定和PCR扩增

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Mercury resistant?Escherichia coli?strains have been isolated from different mercury polluted sites of India and their minimum inhibitory concentration (MIC) levels were determined for HgCl2. Biochemical identification of?E. coli?strains was also performed with HiMedia’s biochemical identification kit for various biochemical assays, like,?Citrate utilization, Lysine decarboxylase, Ornithine decarboxylase, Urease, Phenylalanine deamination, Nitrate reduction, H2S production, Glucose, Adonitol, Lactose, Arabinose, Sorbitol etc.?The zone of inhibition was measured to find the antibiotic susceptibility level. The location of?mer?operon [containing?mergenes including?merA?responsible for the expression of mercuric reductase enzyme (MerA) which converts the toxic Hg2+?to least toxic elemental Hg0] was determined by transforming the isolated plasmids into mercury sensitive host?E. coliDH5α cells. Plasmid isolated from transformed?E. coli?DH5α cells were also analyzed and compared with the plasmid profile of the wild-type?E. coli?strains. Oligonucleotides primer were designed by comparing the known reported sequences of?merA?from Gram-negative bacteria (Escherichia coli?plasmid?R100) and 1695 bp of?merA?gene was amplified by PCR.
机译：从印度不同的汞污染地点分离出了耐汞的大肠杆菌菌株，并确定了它们对HgCl2的最低抑制浓度（MIC）水平。 E的生化鉴定。还使用HiMedia的生化鉴定试剂盒进行了大肠埃希菌菌株的测试，以用于各种生化测定，例如柠檬酸利用，赖氨酸脱羧酶，鸟氨酸脱羧酶，脲酶，苯丙氨酸脱氨，硝酸盐还原，H2S产生，葡萄糖，Adonitol，乳糖，阿拉伯糖，山梨糖醇等。测量抑制区域以发现抗生素敏感性水平。通过将分离的质粒转化为对汞敏感的宿主？确定了“ mer”操纵子的位置（含有负责表达汞还原酶（MerA）的mergenes，merx负责将有毒的Hg2 +α转化为毒性最小的元素Hg0）。 E． coliDH5α细胞。从转化的大肠杆菌分离的质粒。还分析了大肠杆菌DH5α细胞，并与野生型ΔE的质粒图谱进行了比较。大肠杆菌菌株。通过比较革兰氏阴性菌（大肠杆菌质粒pR100）已知的merAβ序列，设计了寡核苷酸引物，并通过PCR扩增了1695bp的merAβ基因。

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