A Novel mRNA Level Subtraction Method for Quick Identification of Target-Orientated Uniquely Expressed Genes Between Peanut Immature Pod and Leaf

Biological Procedures Online

Ning Wu¹  Kanyand Matand¹  Sonya Williams¹ 

[1] Center for Biotechnology Research and Education, Langston University, P.O. Box 1730, Langston, OK 73050, USA

关键词: mRNA;    hybridization;    cDNA library;    gene discovery;    subtraction;    Peanut;   

Others  :  797141
DOI  :  10.1007/s12575-009-9022-z

 received in 2009-07-02, accepted in 2009-11-16,  发布年份 2009

【 摘 要 】

Subtraction technique has been broadly applied for target gene discovery. However, most current protocols apply relative differential subtraction and result in great amount clone mixtures of unique and differentially expressed genes. This makes it more difficult to identify unique or target-orientated expressed genes. In this study, we developed a novel method for subtraction at mRNA level by integrating magnetic particle technology into driver preparation and tester–driver hybridization to facilitate uniquely expressed gene discovery between peanut immature pod and leaf through a single round subtraction. The resulting target clones were further validated through polymerase chain reaction screening using peanut immature pod and leaf cDNA libraries as templates. This study has resulted in identifying several genes expressed uniquely in immature peanut pod. These target genes can be used for future peanut functional genome and genetic engineering research.

【 授权许可】

   
2010 Wu et al; licensee Springer

【 图 表 】

Figure 4.

Figure 2.

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