Cloning and Prokaryotic Expression of Hsp70 from Soybean Cyst Nematode(Heterodera glycines)(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:

2012年02期

Page:

198-202

Research Field:

Publishing date:

Info

Title:

Cloning and Prokaryotic Expression of Hsp70 from Soybean Cyst Nematode(Heterodera glycines)

Author(s):

ZHENG Ya-nan¹; WANG Yuan-yuan²; CHEN Jing-sheng³; ZHU Xiao-feng⁴; CHEN Li-jie⁴; DUAN Yu-xi⁴

1.College of Forestry,Shenyang Agricultural University,Shenyang 110866,Liaoning;

2.College of Bioscience and Biotechnology,Shenyang Agricultural University,Shenyang 110866, Liaoning;

3.Daqing Branch of Heilongjiang Academy of Agricultural Sciences,Daqing 163316,Heilongjiang;

4.College of Plant Protection,Shenyang Agricultural University,Shenyang 110866,Liaoning,China

Keywords:

Heterdara glycines; Heat shock protein 70(Hsp70); Clone; Expression

PACS:

S565.1

DOI:

10.3969/j.issn.1000-9841.2012.02.008

Abstract:

Heat shock protein 70(Hsp70) family is one of the most important heat shock proteins and an evolutionarily conserved protein.It plays an important role in environmental adaptability and stress tolerance of organisms. In this paper,Hsp70 complete cDNA sequence(Hg-Hsp-70)was cloned from Heterodera glycines by RT-PCR and genome walking technology, the full length is 1953 bp,and the Genbank accession number is FJ816100.1.The sequence structure analysis showed that the genomic sequences contained 9 exons and 8 introns,and the sequence homology was higher than other nematodes.This gene encoded 650 amino acids, and the calculated molecular weight was 70 kD,carried two important intact Hsp70 signature sequences.The result of amino acids sequences analysis revealed that translated molecules showed high homology with other nematodes.The prokaryotic expression vector Hsp70pEASY-E1 was constructed for Hg-Hsp-70. Hg-Hsp-70 protein was obviously induced to express by 0.2-1.2 mmol·L^-1 IPTG and reached the maximum expression,after induced 5 h by 0.8 mmol·L^-1 IPTG.

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