Halophiles offer an attractive source of genes conferring salt tolerance. Halobacillus trueperi SS1 strain of Lunsu, Himachal Pradesh, India, a strict halophile, was exploited to isolate and clone the genes for salt tolerance. The genomic library of BamH1 digest of H. trueperi SS1 was constructed in pUC19, and recombinants were screened for salt tolerance on an LB medium containing ampicillin (100 μg/ml) and NaCl (0 to 1.5 M). One recombinant clone named as salt-tolerant clone (STC) conferred salt tolerance to host Escherichia coli/DH5α, which showed growth in the LB medium supplemented with ampicillin and 1.2 M NaCl. Restriction digestion and PCR analysis revealed the presence of an insert of approximately 2000 bp in the STC. DNA sequencing of the 2-kb insert on both strands yielded a sequence of 2301 nucleotides. Protein BLAST analysis of 2301-bp sequence of H. trueperi SS1 present in STC showed 97% identity to multidrug transport ATP binding/permease protein of Halobacillus karajensis. The insert contained in STC was subcloned into pGEX4T2 vector, and the recombinant clone STC/pGEX4T2 conferred salt tolerance to the bacterial host E. coli. The present study led to the isolation of salt tolerance gene encoding a putative multidrug transport ATP binding/permease protein from H. trueperi SS1. The salt tolerance gene can be subcloned for transferring salt tolerance traits into agricultural crop plants for cultivation in saline and coastal lands.

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从印度喜马al尔邦伦苏的嗜盐细菌中分离赋予耐盐性的基因

嗜盐菌提供了赋予耐盐性的诱人基因。利用严格的嗜盐菌，印度喜马al尔邦伦苏的真假单胞菌SS1菌株分离并克隆了耐盐基因。在pUC19中构建真叶假单胞菌SS1的BamH1消化基因组文库，并在含有氨苄青霉素（100μg/ ml）和NaCl（0至1.5 M）的LB培养基上筛选重组子的耐盐性。一个名为耐盐克隆（STC）的重组克隆赋予宿主大肠杆菌/DH5α耐盐性，该克隆在添加氨苄青霉素和1.2 M NaCl的LB培养基中显示出生长。限制性酶切和PCR分析显示STC中存在大约2000bp的插入片段。对两条链上的2-kb插入片段进行DNA测序得到了2301个核苷酸的序列。在STC中存在的Trueperi SS1的2301 bp序列的蛋白质BLAST分析显示，与哈洛杆菌（Halobacillus karajensis）的多药转运ATP结合/渗透酶蛋白具有97％的同一性。将STC中包含的插入片段亚克隆到pGEX4T2载体中，重组克隆STC / pGEX4T2使细菌宿主大肠杆菌具有耐盐性。本研究导致了从H.trueperi SS1分离出一种编码耐盐基因的基因，该基因编码一种假定的多药转运ATP结合/渗透酶蛋白。可以将耐盐基因亚克隆，以将耐盐性状转移到农作物中，以在盐碱地和沿海地区种植。重组克隆STC / pGEX4T2赋予细菌宿主大肠杆菌耐盐性。本研究导致了从H.trueperi SS1分离出一种编码耐盐基因的基因，该基因编码一种假定的多药转运ATP结合/渗透酶蛋白。可以将耐盐基因亚克隆，以将耐盐性状转移到农作物中，以在盐碱地和沿海地区种植。重组克隆STC / pGEX4T2赋予细菌宿主大肠杆菌耐盐性。本研究导致了从H.trueperi SS1分离出一种编码耐盐基因的基因，该基因编码一种假定的多药转运ATP结合/渗透酶蛋白。可以将耐盐基因亚克隆，以将耐盐性状转移到农作物中，以在盐碱地和沿海地区种植。