Nagamani Gorantla, Alex Swapna, Soni K B, Anith K N, Viji M M, Kiran A G
Department of Plant Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram, Kerala 695522 India.
Department of Agricultural Microbiology, College of Agriculture, Vellayani, Thiruvananthapuram, Kerala 695522 India.
3 Biotech. 2019 Mar;9(3):113. doi: 10.1007/s13205-019-1640-9. Epub 2019 Mar 1.
The present study is the first report on the application of silver nanoparticles for efficient bacterial transformation. EC50 value of 100 nm silver nanoparticles against DH5α cells was recorded as 4.49 mg L in toxicity assay. Competency induction in DH5α cells by treatment with 100 nm silver nanoparticles at a concentration of 1 mg L for 60 min and transformation using three plasmid vectors of different sizes, viz. pUC18, pBR322 and pCAMBIA resulted in tenfold increase in the bacterial transformation efficiency, i.e. 8.3 × 10, 8.0 × 10 and 7.9 × 10 cfu ng of DNA, respectively, even without heat shock compared to the conventional chemical method using 0.1 M calcium chloride (2.3 × 10 cfu ng of DNA).
本研究是关于银纳米颗粒在高效细菌转化中应用的首次报道。在毒性试验中,100纳米银纳米颗粒对DH5α细胞的半数有效浓度(EC50)记录为4.49毫克/升。用浓度为1毫克/升的100纳米银纳米颗粒处理DH5α细胞60分钟进行感受态诱导,并使用三种不同大小的质粒载体,即pUC18、pBR322和pCAMBIA进行转化,即使不进行热休克处理,细菌转化效率也比使用0.1M氯化钙的传统化学方法(2.3×10菌落形成单位/纳克DNA)提高了十倍,分别为8.3×10、8.0×10和7.9×10菌落形成单位/纳克DNA。