抵抗性
生物
底漆(化妆品)
DNA微阵列
炸薯条
计算生物学
环境化学
生化工程
基因
遗传学
化学
计算机科学
基因组
基因表达
工程类
电信
流动遗传元素
有机化学
作者
Jiaojiao Zhu,Qiwei Huang,Peng Xu,Xinyuan Zhou,Shenghan Gao,Yuanping Li,Xuesong Luo,Yi Zhao,Christopher Rensing,Jian‐Qiang Su,Peng Cai,Yurong Liu,Wenli Chen,Xiuli Hao,Qiaoyun Huang
标识
DOI:10.1021/acs.est.2c00488
摘要
Bacterial metal detoxification mechanisms have been well studied for centuries in pure culture systems. However, profiling metal resistance determinants at the community level is still a challenge due to the lack of comprehensive and reliable quantification tools. Here, a novel high-throughput quantitative polymerase chain reaction (HT-qPCR) chip, termed the metal resistance gene (MRG) chip, has been developed for the quantification of genes involved in the homeostasis of 9 metals. The MRG chip contains 77 newly designed degenerate primer sets and 9 published primer sets covering 56 metal resistance genes. Computational evaluation of the taxonomic coverage indicated that the MRG chip had a broad coverage matching 2 kingdoms, 29 phyla, 64 classes, 130 orders, 226 families, and 382 genera. Temperature gradient PCR and HT-qPCR verified that 57 °C was the optimal annealing temperature, with amplification efficiencies of over 94% primer sets achieving 80-110%, with R2 > 0.993. Both computational evaluation and the melting curve analysis of HT-qPCR validated a high specificity. The MRG chip has been successfully applied to characterize the distribution of diverse metal resistance determinants in natural and human-related environments, confirming its wide scope of application. Collectively, the MRG chip is a powerful and efficient high-throughput quantification tool for exploring the microbial metal resistome.
科研通智能强力驱动
Strongly Powered by AbleSci AI