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一种用于测定微量样品中转基因溶菌酶山羊含量的事件特异性实时荧光定量PCR方法。

An Event-Specific Real-Time PCR Method for Measuring Transgenic Lysozyme Goat Content in Trace Samples.

作者信息

Xu Wenting, Cui Jinjie, Liu Biao, Yang Litao

机构信息

Joint International Research Laboratory, Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.

State Key Laboratory, Cotton Biology, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang 455000, China.

出版信息

Foods. 2021 Apr 23;10(5):925. doi: 10.3390/foods10050925.

Abstract

Lysozymes are used in sterilisation, antisepsis, dairy additives, inflammation, and cancer. One transgenic goat line expressing high levels of human lysozyme (hLZ) in goat milk has been developed in China. Herein, we established an event-specific real-time polymerase chain reaction (real-time PCR) method to detect the transgenic hLZ goat line. The developed method has high specificity, sensitivity and accuracy, and a wide quantitative dynamic range. The limit of detection and limit of quantification was 5 and 10 copies per reaction, respectively. The practical sample analysis results showed that the method could identify and quantify transgenic lysozyme content in trace samples in routine lab analyses. Furthermore, the potential applicability in risk assessment, such as molecular characterisation and gene horizontal transfer, was confirmed. We believe that this method is suitable for the detection of transgenic hLZ goat line and its derivate.

摘要

溶菌酶用于杀菌、防腐、乳制品添加剂、炎症和癌症治疗。中国已培育出一种在羊奶中高水平表达人溶菌酶(hLZ)的转基因山羊品系。在此,我们建立了一种事件特异性实时聚合酶链反应(实时PCR)方法来检测转基因hLZ山羊品系。所建立的方法具有高特异性、高灵敏度和高准确性,并且定量动态范围宽。检测限和定量限分别为每个反应5拷贝和10拷贝。实际样品分析结果表明,该方法能够在常规实验室分析中识别和定量痕量样品中转基因溶菌酶的含量。此外,还证实了该方法在风险评估中的潜在适用性,如分子特征分析和基因水平转移。我们认为该方法适用于转基因hLZ山羊品系及其衍生物的检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e251/8146569/97b1cb068cca/foods-10-00925-g001.jpg

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