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《Science of Sericulture》 2007-01
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Cloning and Sequence Analysis of cDNA of Diapause Biological Clock Protein Esterase A4 of Bombyx mori

WANG Yu-Jun XU Shi-Qing* SIMA Yang-Hu WU Yang-Chun LIU YingLIU Xue-Guang CONG Hai-Feng(College of Life Science,Soochow University,Suzhou Jiangsu 215123,China)  
The diapause biological clock protein of Bombyx silkworm is a count down clock which depends on esterase A4(EA4) in their eggs.EA4 was the first confirmed time interval measuring enzyme(TIME).A 308 bp of ea4 cDNA sequence was generated by RT-PCR with degenerate primers designed according to the determined amino acids and using the total RNA of fat body and ovary from 3-day old female pupa as the template.The consistence of 1-144 nt and 145-308 nt of the sequence with 5 595-5 738 nt of whole genome shotgun sequence(WGS)(AADK01019126.1) and 8 636-8 473 nt of WGS(AADK01014467.1) were 95% and 98%,respectively.According to the sequence of the ea4 gene fragment obtained,ea4 gene integrity cDNA sequence was forecasted by method of bioinformatics and the genome data of Bombyx mori.Then,a pair of special primers was designed according to the forecasted cDNA sequence of ea4,and the complete cDNA sequence was obtained by RT-PCR using the total RNA of fat body and ovary of female pupa as the template.The cDNA of ea4 gene was 519 bp in length,in which 1-48 bp was the signal peptide sequence.And the amino acid sequence encoded by 49-519 bp showed 98% similarity with that of EA4 protein,with 3 amino acids difference at the C-terminal,in which the 156th was the first defined one.
【Fund】: 国家自然科学基金项目(编号30371086);; 国家重点基础研究发展计划“973”项目(编号2005CB121005)
【CateGory Index】: S881.26
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