|本期目录/Table of Contents|

[1]孙恒文△,胡义德,曾子君,等.人线粒体DNA基因表达谱芯片的制备及鉴定[J].生物医学工程研究,2015,03:164-169.
 SUN Hengwen,HU Yide,ZENG Zijun,et al.Manufacture and Identification of Human mtDNA Genes Expressing Microarray[J].Journal of Biomedical Engineering Research,2015,03:164-169.
点击复制

人线粒体DNA基因表达谱芯片的制备及鉴定(PDF)

《生物医学工程研究》[ISSN:1006-6977/CN:61-1281/TN]

期数:
2015年03期
页码:
164-169
栏目:
出版日期:
2015-09-25

文章信息/Info

Title:
Manufacture and Identification of Human mtDNA Genes Expressing Microarray
文章编号:
1672-6278 (2015)03-0164-06
作者:
孙恒文1△胡义德2曾子君1潘燚1方良毅1谭佩欣1曾向伟3
1.广东省人民医院(广东省医学科学院)肿瘤中心放疗科,广州 510080;2.第三军医大学附属新桥医院全军肿瘤中心,重庆 400037;3.广东医学院附属彭湃纪念医院肿瘤科,广东 海丰 516400
Author(s):
SUN Hengwen1 HU Yide2 ZENG Zijun1 PAN Yi1 FANG Liangyi1 TAN Peixin1 ZENG Xiangwei3
1.Department of Radiation, Cancer Center of Guangdong General Hospital (Guangdong Academy of Medical Science), Guangzhou 510080, China;2.Cancer Center of PLA, Xinqiao Hospital, Third Military Medical University, Chongqing 400037,China;3.Department of Oncology, Guangdong Medical College Affiliated Pengpai Memorial Hospital, Haifeng 516400, Guangdong, China
关键词:
线粒体DNA基因表达寡核苷酸芯片信使RNA核糖体RNA转运RNA
Keywords:
Mitochondrial DNA Gene expression Oligonuleotide microarray mRNA rRNA tRNA
分类号:
R378
DOI:
-
文献标识码:
A
摘要:
研制人线粒体DNA(mitochondrial DNA,mtDNA)表达谱芯片,鉴定芯片的特异性和稳定性,完善芯片的制备和使用程序,优化实验参数,为线粒体功能研究提供物质基础及技术保障。以人mtDNA剑桥序列为标准,设计mtDNA编码的13种mRNA、2种rRNA和9种tRNA基因及蛋白产物定位于线粒体的核DNA(nDNA)编码的5种凋亡相关基因寡核苷酸探针。芯片点样仪点制芯片,进行荧光显示和洗脱,用其检测人宫颈癌上皮Hela细胞和人肾小管上皮Hc1细胞cDNA文库mtDNA编码基因及nDNA编码凋亡相关基因的差异表达情况。所点制的芯片扫描结果显示,样点分布均匀、清晰,规整度好,无漏点、连点。cDNA文库杂交鉴定结果显示,整张芯片荧光信号均匀一致,背景清晰,各基因重复样点杂交结果一致,各质控点能正常显示。成功制备了人mtDNA基因表达谱芯片,完善了该芯片的制备和使用程序,通过初步应用,证明所研制的人mtDNA基因表达谱芯片具有特异性高、稳定性好等优点,可用于不同组织或细胞样本cDNA文库mtDNA基因的差异表达分析。
Abstract:
To manufacture and identificate the expressing microarray of human mtDNA genes. In the process of conformation, the specificity and stability of the new microarray were evaluated, the procedure of preparing and applying of the new microarray were consummated, and the experimental parameter were optimised to provide the substance support and technology guarantee for the function research of mitochondrial genome. Design of the oligonucleotide probes of 13 mRNA、2 rRNA、9 tRNA encoded by mtDNA was based on the mtDNA Cambridge Sequence, and the oligonucleotide probes of 5 apoptosis related genes encoded by nuclear genome was based on the database of Genebank. The microarray was printed with microarray robots. Fluorescence background manifest and deletion were according to the standard process. The primary microarray was firstly used to detect the differential expressions of mtDNA genes and apoptosis related genes between cDNA library of human cervix epithelia carcinoma cell line Hela and human renal tubule epithelia cell line Hc1. The printed spots on the primary microarray show distinct and regular on the scanned imagine, and the background of the slides is nearly undetected. The hybridized imagine of cDNA library from cell lines show a good quality in size and color contrast. Twenty mini-blocks are printed in the first round.

参考文献/References

备注/Memo

备注/Memo:
(收稿日期:2015-03-26)国家自然科学基金资助项目(81302033)通信作者Email:sunrise761114@foxmail.com
更新日期/Last Update: 2016-07-25