Identification of a Bombyx mori gene encoding small heat shock protein BmHsp27.4 expressed in response to high-temperature stress.
Elucidating the mechanisms underlying the response and resistance to high-temperature stress in the Lepidoptera is essential for understanding the effect of high-temperature on the regulation of gene expression. A tag (CATGAACGTGAAGAGATTCAG) matching the predicted gene BGIBMGA005823-TA in SilkDB identified the most significant response to high-temperature stress in a screen of the heat-treated digital gene expression library of Bombyx mori (B. mori) (Unpublished data). BLAST and RACE showed that the gene is located on chromosome 5 and has an open reading frame (ORF) of 741bp. Phylogenetic analysis found that B. mori small heat shock protein 27.4 (BmHSP27.4) is in an evolutionary branch separate from other small heat shock proteins. Expression analysis showed that BmHsp27.4 is highly expressed in brain, eyes and fat bodies in B. mori. Its mRNA level was elevated at high-temperature and this increase was greater in females. The ORF without the signal peptide sequence was cloned into vector pET-28a(+), transformed and over-expressed in Escherichia coli Rosetta (DE3). Western blotting and immunofluorescence analysis with a polyclonal antibody, confirmed that the level of protein BmHSP27.4 increased at a high-temperature, in accordance with its increased mRNA level. In this study, BmHsp27.4 was identified as a novel B. mori gene with an important role in response to high-temperature stress.
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Hua
H
Wang
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Yan
Y
Fang
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Zhongzan
Z
Bao
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Xing
X
Jin
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Wenjuan
W
Zhu
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Lipeng
L
Wang
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Teng
T
Liu
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Haipeng
H
Ji
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China.
Haiying
H
Wang
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China; National Engineering Laboratory for Modern Silk, Soochow University, China.
Shiqing
S
Xu
Department of Applied Biology, School of Biology and Basic Medical Sciences, Medical College of Soochow University, China; National Engineering Laboratory for Modern Silk, Soochow University, China. Electronic address: simyh@suda.edu.cn.
Yanghu
Y
Sima
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D005260:Female; D055551:HSP27 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018869:Heat-Shock Response; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012737:Sex Factors
D055551:HSP27 Heat-Shock Proteins; D019476:Insect Proteins; D012333:RNA, Messenger
Bombyx mori; Immunofluorescence; Prokaryotic expression; Small heat shock protein