The microbiota inhabiting in metal polluted environment develops strong defense mechanisms to combat pollution and sustain life. Investigating the functional genes of the eukaryotic microbiota inhabiting in these environments by using metatranscriptomics approach was the focus of this study. Size fractionated eukaryotic cDNA libraries (library A, < 0.5 kb, library B, 0.5-1.0 kb, and library C, > 1.0 kb) were constructed from RNA isolated from the metal contaminated soil. The library C was screened for Cadmium (Cd) tolerant genes by using Cd sensitive yeast mutant ycf1^Δ by functional complementation assay, which yielded various clones capable of growing in Cd amended media. One of the Cd tolerant clones, PLCg39 was selected because of its ability to grow at high concentrations of Cd. Sequence analysis of PLCg39 showed homology with DHHC palmitoyl transferases, which are responsible for addition of palmitoyl groups to proteins and usually possess metal coordination domains. The cDNA PLCg39 was able to confer tolerance to Cd-sensitive (ycf1^Δ), Copper-sensitive (cup1^Δ) and Zn-sensitive (zrc1^Δ) yeast mutants when grown at different concentrations of Cd (40-100 μM), Cu (150-1000 μM) and Zn (10-13 mM), respectively. The DHHC mutant akr1^Δ transformed with PLCg39 rescued from the metal sensitivity indicating the role of DHHC palmitoyl transferase in metal tolerance. This study demonstrated that PLCg39 acts as a potential metal tolerant gene which could be used in bioremediation, biosensing and other biotechnological fields.

居住在金属污染环境中的微生物群发展了强大的防御机制，以抗击污染并维持生命。本研究的重点是通过元转录组学方法研究居住在这些环境中的真核微生物群的功能基因。从分离自金属污染土壤的RNA中构建大小分级的真核cDNA文库（文库A，<0.5 kb，文库B，0.5-1.0 kb，文库C，> 1.0 kb）。程序库C是通过使用镉敏感突变体酵母筛选镉（Cd）耐受性基因ycf1 ^Δ通过功能互补分析，得到了能够在Cd改良培养基中生长的各种克隆。选择了一种耐Cd的克隆PLCg39，因为它具有在高Cd浓度下生长的能力。PLCg39的序列分析表明与DHHC棕榈酰基转移酶具有同源性，后者负责向蛋白质中添加棕榈酰基，通常具有金属配位域。将cDNA PLCg39能够赋予对镉敏感（ycf1 ^Δ），铜-敏感（CUP1 ^Δ）和Zn-敏感（zrc1 ^Δ）酵母突变体当在不同浓度的镉（40-100微米），铜（的生长150-1000μM）和Zn（10-13 mM）。所述突变体DHHC akr1 ^Δ从金属敏感性中拯救了用PLCg39转化的DNA，表明DHHC棕榈酰转移酶在金属耐受性中的作用。这项研究表明，PLCg39是潜在的金属耐受基因，可用于生物修复，生物传感和其他生物技术领域。