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Title: <span style="font-size:11.0pt;font-family: "Times New Roman";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN-GB">Cloning and expression of a small heat and salt tolerant protein (<i>Hsp</i>22) from <i>Chaetomium globosum</i></span>
Authors: Aggarwal, Rashmi
Gupta, Sangeeta
Sharma, Sapna
Banerjee, Sagar
Singh, Priyanka
Keywords: <i>Chaetomium globosum</i>
Gene cloning
Small heat shock protein
Stress tolerance
Issue Date: Nov-2012
Publisher: NISCAIR-CSIR, India
Abstract:   The present study reports molecular characterization of small heat shock protein gene in Indian isolates of <i style="mso-bidi-font-style:normal">Chaetomium globosum</i>, <i style="mso-bidi-font-style:normal">C. perlucidum, C. reflexum, C. cochlioides </i>and<i style="mso-bidi-font-style:normal"> C. cupreum. </i>Six isolates of <i style="mso-bidi-font-style:normal">C. globosum</i> and other species showed a band of 630bp using specific primers. Amplified cDNA product of <i style="mso-bidi-font-style:normal">C. globosum</i> (Cg 1) cloned and sequenced showed 603bp open reading frame encoding 200 amino-acids. The protein sequence had a molecular mass of 22 kDa and was therefore, named Hsp22. BlastX analysis revealed that the gene codes for a protein homologous to previously characterized <i style="mso-bidi-font-style:normal">Hsp</i>22.4 gene from <i>C. globosum</i><span style="mso-bidi-font-style:italic"> (<span style="mso-fareast-font-family:Calibri;letter-spacing:-.1pt" lang="EN-GB">AAR36902.1, XP 001229241.1) <span style="letter-spacing:-.1pt;mso-bidi-font-style: italic" lang="EN-GB">and shared 95% identity in amino acid sequence. It also showed varying degree of similarities with small Hsp protein from <i>Neurospora</i> spp. (60%), <i style="mso-bidi-font-style: normal">Myceliophthora</i> sp. (59%), <i style="mso-bidi-font-style:normal">Glomerella</i> sp. (50%), <i style="mso-bidi-font-style:normal">Hypocrea</i> sp. (52%), and <i style="mso-bidi-font-style:normal">Fusarium</i> spp. (51%). This gene was further cloned into pET28a (+) and transformed <i>E. coli</i><span style="mso-bidi-font-style:italic"> BL21 cells were induced by IPTG, and the expressed protein of 30 kDa was analyzed by SDS-PAGE. <span style="mso-bidi-font-style:italic">The IPTG induced transformants displayed significantly greater resistance to NaCl and Na<sub>2</sub>CO<sub>3</sub> stresses. </span></span></span></span></span>
Description: 826-832
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.50(11) [November 2012]

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