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中国生物工程杂志 2015.5 Vol 35, No.5     
基因差异转录表达验证方法研究进展
 
左海洋, 陈晓丽, 蔡勇, 郝海生, 秦彤, 赵学明, 路永强, 王栋
1. 吉林农业大学动物科学技术学院 长春 130118
 
Key Words: 基因差异表达 荧光定量PCR 核酸杂交 核酸探针
 
 
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Abstract

基因的差异表达与组织、细胞的生物学性状和功能密切相关,随着对组织、器官分化及细胞、生物个体生长发育等的深入研究,大量差异转录基因的验证分析变得越来越重要。为此,该领域基于核酸互补配对和PCR原理建立了Northern blot和荧光定量PCR等一系列基因差异转录分析技术,利用这些方法对不同处理、不同组织器官、不同发育时段的基因差异转录进行了验证分析,为后续的基因功能分析奠定了坚实基础,并且通过检测分析,使基因差异表达分析方法由定性到定量、由繁琐复杂到简单快速、由以大量RNA为前提到对少量RNA样品的检测,甚至建立了单细胞荧光定量PCR方法,基因差异转录验证方法正在向更高效、精准方向发展,并使成本逐步降低。但是,到目前为止,生殖细胞等样品取样较为困难、精子等RNA含量较少样品的基因差异表达验证挑战性还仍然很大。从验证方法发展历程的角度,对基因差异转录验证方法进行了总结,希望为取样难度较大、RNA含量较低样品的基因差异表达分析提供有价值的参考。
 
Abstract

The differential expression of genes is closely related to biological characters and function of tissue and cell. With the extensive study on the differentiation of tissue and organs and the growth and development of cells and individuals, the validation of differentially expressed genes are getting more and more important. Thus, a series of validating techniques of differentially expressed analysis were established on the basis of nucleic acid hybridization and PCR, such as Northern blot and fluorescence quantitative PCR. The differentially expressed genes were validated of different treatment, different organs and different development period using these methods, which laid a solid foundation for the gene functional analysis. With the fast development of the detecting methods, the validation methods of differential expression of genes has developed from qualitative to quantitative, from tedious and complicated to simple and rapid, from requiring a large number of initial RNA to only a small amount of RNA, even the fluoresce quantitative PCR method for single cell has been established, and the method is to be more efficient, accurate and low cost. However, The validation for the samples difficult sampling like germ cell and less amount of RNA containing like sperm is still a great challenge for the current method. The validation methods of differential expression of genes from the perspective of the development process was reviewed. Reference to the detedtion of samples dificult sampling and less amount RNA containing was provided.
 
 
 
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中国科学院遗传与发育生物学研究所
中国科学院遗
中国科学院遗传与发育生物学研究所 中国遗传学会
中国科学院遗传与发育生物学
中国科


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