Identification of Differential Genes by Suppression Subtractive Hybridization: V. PCR-Based DNA Dot Blot

2008 ◽  
Vol 2008 (8) ◽  
pp. pdb.prot4859-pdb.prot4859
Author(s):  
D. V. Rebrikov
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Subtractive Hybridization ◽  
Dot Blot ◽  
Differential Genes

Identification and Analysis of Genes Present in Leptospira interrogans serovar lai but Absent in L. biflexa serovar monvalerio

2004 ◽  
Vol 36 (12) ◽  
pp. 832-839 ◽  
Author(s):  
Ping He ◽  
Xiang-Yan Zhang ◽  
Xiao-Kui Guo ◽  
Bao-Yu Hu ◽  
Xiao-Tian Huang ◽  
...  
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Dna Sequences ◽  
Subtractive Hybridization ◽  
Leptospira Interrogans ◽  
Bacterial Strains ◽  
Dot Blot ◽  
Protein Coding ◽  
Domain Structures ◽  
Genes Encoding ◽  
Close Relatives

Abstract Genes present in virulent bacterial strains but absent in avirulent close relatives can be of great biologic and clinical interest. This project aimed to identify strain specific DNA sequences of Leptospira interrogans serovar lai, which is absent in the saprophytic L. biflexa serovar monvalerio, via suppression subtractive hybridization with the former as the tester while the latter as the driver. The mixture of PCR amplified DNA fragments from two subtractive hybridization experiments were cloned into pMD18-T vector and the positive clones were identified by dot blotting against the chromosome DNA of the two strains individually. After DNA sequencing and analysis, the distribution of these genomic fragment sequences in a panel of pathogenic and nonpathogenic leptospires was investigated employing dot blot analysis. Among the 188 positive clones randomly chosen, 24 contained the tester strain specific genomic regions, of which, 5 were non-coding fragments while the others contained 23 distinct protein coding sequences. Besides 9 genes encoding functional proteins, 12 genes encode unknown proteins and the rest two genes encode proteins with recognizable domain structures, one for a putative leucine-rich repeats (LRR) family protein while the other as an outer-membrane protein. Our experiment results indicated that suppression subtractive hybridization is effective for screening specific DNA sequences between two leptospiral strains, and some of these sequences might be responsible for virulence determination. Further analysis of these DNA sequences will provide important information on the pathogenesis of Leptospira.

Identification of Differential Genes by Suppression Subtractive Hybridization: An Overview

2008 ◽  
Vol 2008 (8) ◽  
pp. pdb.top21-pdb.top21 ◽  
Author(s):  
D. V. Rebrikov
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Subtractive Hybridization ◽  
Differential Genes

Construction and analysis of gonad suppression subtractive hybridization library of rice field eel (Monopterus albus)

2013 ◽  
Vol 18 (1) ◽  
pp. 23-28
Author(s):  
Xiancheng QU ◽  
Xiaoli SHANG ◽  
Cui CHENG ◽  
Xuewei QU ◽  
Yong ZHANG ◽  
...  
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Rice Field ◽  
Subtractive Hybridization ◽  
Suppression Subtractive Hybridization Library ◽  
Monopterus Albus ◽  
Rice Field Eel
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