Real-Time Quantitative PCR Analysis of the Expression Pattern of the Hypoglycemic Polypeptide-P Gene in Momordica charantia.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS

ACS Applied Bio Materials Pub Date : 2019-12-16 DOI:10.3390/genes10121044

Yi-Shuai Wang, Xiang-Qing Zeng, Xu-Zhong Yang, Wei Liu, Peng-Fei Li, Fu-Jun Wang, Jian Zhao

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Abstract

This study was designed to establish a real-time quantitative polymerase chain reaction (qPCR) method to rapidly and reliably analyze the hypoglycemic polypeptide-P gene expression pattern in Momordica charantia (MC) and to examine its expression changes in different MC accessions, harvesting seasons, and tissue types. The qPCR results were further verified by using Western blotting (WB). A total of 10 MCs with different accessions were collected and tested in this study. Among the tested accessions, RU5H showed the highest expression level of the polypeptide-P gene. The expression level of the polypeptide-P gene was not only season-related (with the highest in early July) but also tissue-related (with the highest in the seed tissue). In addition, the expression characteristic of the polypeptide-P gene was maturity-related, with the highest expression level in the tender MC. The WB results show that the transcription level of this gene shows an almost similar trend to the corresponding protein expression level. In conclusion, the established qPCR method can rapidly and effectively detect the expression levels of the polypeptide-P gene in MCs with different accessions; furthermore, various factors, including the accessions, harvesting seasons, and tissue types can affect the expression level.

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苦瓜降血糖多肽- p基因表达模式的实时定量PCR分析。

本研究旨在建立一种实时定量聚合酶链反应(qPCR)方法，快速、可靠地分析苦瓜(Momordica charantia, MC)降血糖多肽- p基因的表达谱，并检测其在不同MC品种、收获季节和组织类型中的表达变化。采用Western blotting (WB)进一步验证qPCR结果。本研究共收集了10份不同加入量的MCs，并对其进行了测试。在供试品种中，RU5H多肽- p基因表达量最高。多肽- p基因的表达量不仅与季节有关(7月初最高)，而且与组织有关(种子组织中最高)。此外，多肽- p基因的表达特征与成熟度相关，在嫩嫩MC中表达量最高。WB结果显示，该基因的转录水平与相应的蛋白表达水平呈现几乎相似的趋势。综上所述，所建立的qPCR方法能够快速有效地检测不同加入量MCs中多肽- p基因的表达水平;此外，品种、收获季节和组织类型等多种因素都会影响其表达水平。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ACS Applied Bio Materials Chemistry-Chemistry (all)

CiteScore

9.40

自引率

2.10%

发文量

464

期刊介绍： ACS Applied Bio Materials is an interdisciplinary journal publishing original research covering all aspects of biomaterials and biointerfaces including and beyond the traditional biosensing, biomedical and therapeutic applications. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrates knowledge in the areas of materials, engineering, physics, bioscience, and chemistry into important bio applications. The journal is specifically interested in work that addresses the relationship between structure and function and assesses the stability and degradation of materials under relevant environmental and biological conditions.