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基于量子点标记的新型核酸探针原位杂交方法

发表时间:2013-06-15  浏览量:1126  下载量:340
全部作者: 贺佳玮,邹铅,刘军安,王旭,曹建华,赵书红
作者单位: 华中农业大学动物科技学院,农业动物遗传育种与繁殖教育部重点实验室&农业部猪遗传育种重点开放实验室;华中农业大学理学院;华中农业大学动物医学院,国家兽药残留基准实验室
摘 要: 量子点(quantum dot,QD)具有荧光稳定性强、检测灵敏度高、光谱宽和生物相容性好等诸多优点。研究通过化学合成CdSe/ZnS核壳型QD核心,共价结合靶基因核酸片段形成QD连接的甘油醛-3-磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase, GAPDH)基因探针,经电泳检测及荧光表征,证实该探针具有QD和核酸探针双重特性。在原位杂交实验中,使用该新型QD-核酸探针对大白母猪背最长肌石蜡切片中的靶基因进行检测,成功检测到阳性信号。研究为基于QD标记的核酸探针原位杂交实验提供了实验基础,为QD这一新型荧光材料在生物领域的应用提供了依据,同时为原位杂交应用于育种实践提供了可行的方法。
关 键 词: 家畜育种学;量子点;甘油醛-3-磷酸脱氢酶基因;核酸探针;原位杂交
Title: In situ hybridization protocol using a new type of nucleic acid probe based on quantum dots
Author: HE Jiawei, ZOU Qian, LIU Jun’an, WANG Xu, CAO Jianhua, ZHAO Shuhong
Organization: Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture, College of Animal Science, Huazhong Agricultural University; College of Science, Huazhong Agricultural Universtiy; National Reference Laboratory of Veterinary Drug Residues, College of Veterinary Medicine, Huazhong Agricultural University
Abstract: Quantum dot (QD) is a new material in biomarkers, which has significant advantages of stable fluorescence, high sensitivity, wide fluorescence emission spectrum and harmonious biocompatibility. In this study, the cores of CdSe/ZnS-QDs was chemically synthesized, and then they were covalently binded with nucleic acid probes from glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene to form QD-DNA probes. This probe was proved to have the characteristics of QD and nucleic acid probe by electrophoresis and fluorescence detection. Furthermore, the QD-DNA probes were used in the in situ hybridization to conduct for the target gene detection in longissimus dorsi muscle in Yorkshire pigs. The results showed that QD-labeled probe had distinct positive signals. These results provided an experimental foundation for in situ hybridization based on QD-labeled probes, and shed light on the biomaterial applications of QDs. In addition, this research also provided a feasible method for the application of in situ hybridization in animal breeding.
Key words: animal breeding; quantum dot; glyceraldehyde-3-phosphate dehydrogenase gene; DNA probe; in situ hybridization
发表期数: 2013年6月第11期
引用格式: 贺佳玮,邹铅,刘军安,等. 基于量子点标记的新型核酸探针原位杂交方法[J]. 中国科技论文在线精品论文,2013,6(11):1064-1069.
 
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