NDLI: Construction of cold induced subtracted cDNA library from Cicer microphyllum and transcript characterization of identified novel wound induced gene

Content Provider	Springer Nature Link
Author	Singh, Rupesh Kumar Singh, Shweta Pandey, Pankaj Anandhan, Sivalingam Goyary, Danswrang Pande, Veena Ahmed, Zakwan
Copyright Year	2012
Abstract	A forward cold induced subtracted cDNA library was constructed to identify the stress regulated genes in a high altitude cold desert-adapted species Cicer microphyllum, a wild relative of cultivated chickpea, distributed in western and trans-Himalayas. A total of 1,040 clones were obtained from the subtracted cDNA library. These clones were screened for presence of insert with colony PCR. A total of 523 clones were picked by colony PCR among which 300 clones were observed differentially expressed as per dot blot analysis. Differentially expressed clones were sequenced and assembled into clusters based on the presence of overlapping, identical, or similar sequences. A total of 283 ESTs were submitted in gene bank (accession numbers GO241043 to GO241326). BLAST analysis of these ESTs revealed its similarity for regulatory proteins like kinases, metallothionin, and enzymes/proteins with unknown functions. A cDNA encoding wound induced like protein, identified from this cold induced subtraction cDNA library, was full-length cloned using RACE and sequenced (accession number GQ914056). Southern blot of C. microphyllum indicated single copy of the gene in genome. Transcript expression profiling of this gene by quantitative real-time PCR and northern blot confirmed its up-regulation during low temperature stress. Further, in situ RNA hybridization also revealed cold (4°C) induced expression of the gene.
Starting Page	459
Ending Page	469
Page Count	11
File Format	PDF
ISSN	0033183X
Journal	Protoplasma
Volume Number	250
Issue Number	2
e-ISSN	16156102
Language	English
Publisher	Springer Vienna
Publisher Date	2012-07-15
Publisher Place	Vienna
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Cicer microphyllum Low temperature Suppression subtraction hybridization (SSH) Wound induced gene qRT PCR In situ hybridization Cell Biology Plant Sciences Zoology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Plant Science Medicine

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