Arid
DOI10.1007/s00709-012-0429-z
Construction of cold induced subtracted cDNA library from Cicer microphyllum and transcript characterization of identified novel wound induced gene
Singh, Rupesh Kumar1; Singh, Shweta1; Pandey, Pankaj1; Anandhan, Sivalingam1; Goyary, Danswrang1; Pande, Veena2; Ahmed, Zakwan1
通讯作者Goyary, Danswrang
来源期刊PROTOPLASMA
ISSN0033-183X
EISSN1615-6102
出版年2013
卷号250期号:2页码:459-469
英文摘要

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 (4A degrees C) induced expression of the gene.


英文关键词Cicer microphyllum Low temperature Suppression subtraction hybridization (SSH) Wound induced gene qRT PCR In situ hybridization
类型Article
语种英语
国家India
收录类别SCI-E
WOS记录号WOS:000316572400005
WOS关键词SALICYLIC-ACID ; ABSCISIC-ACID ; CLP PROTEASE ; ARABIDOPSIS ; STRESS ; PROTEINASE ; EXPRESSION ; ACCLIMATION ; TOLERANCE ; RESISTANCE
WOS类目Plant Sciences ; Cell Biology
WOS研究方向Plant Sciences ; Cell Biology
资源类型期刊论文
条目标识符http://119.78.100.177/qdio/handle/2XILL650/179521
作者单位1.Minist Def, Def Inst Bioenergy Res, Haldwani 263139, Uttarakhand, India;
2.Kumaon Univ, Dept Biotechnol, Naini Tal 263001, Uttarakhand, India
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GB/T 7714
Singh, Rupesh Kumar,Singh, Shweta,Pandey, Pankaj,et al. Construction of cold induced subtracted cDNA library from Cicer microphyllum and transcript characterization of identified novel wound induced gene[J],2013,250(2):459-469.
APA Singh, Rupesh Kumar.,Singh, Shweta.,Pandey, Pankaj.,Anandhan, Sivalingam.,Goyary, Danswrang.,...&Ahmed, Zakwan.(2013).Construction of cold induced subtracted cDNA library from Cicer microphyllum and transcript characterization of identified novel wound induced gene.PROTOPLASMA,250(2),459-469.
MLA Singh, Rupesh Kumar,et al."Construction of cold induced subtracted cDNA library from Cicer microphyllum and transcript characterization of identified novel wound induced gene".PROTOPLASMA 250.2(2013):459-469.
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