Cloning Of The Heat Shock Protein Gene From The Diamondback Moth, Plutella Xylostella And The Expression Of MRNA

This paper maked use of the related knowledge of molecular biology to clone the segment of hsp70 gene of Diamondback Moth under the condition of our lab study about the relationship of the fitness and the HSP70 expression quantity of drug-fast Diamondback Moth and drug-sensitive Diamondback Moth at 37℃. We taked this part as the template to synthesize a probe to carry on Northern Blot in order to study the different of the hsp70 mRNA expression quantity of them. We landed Genbank , blasted many hsp70 mRNAs of insect and select the conservative district to design two pairs of prime. The purification of PCR , after carrying on the TA clone, sended Invitrogen company to measure the sequence.The main results as followings:1. Two segments, 600bp and 680bp,have been gotten. the homology was between83% and 86% after carrying on the BLAST with orther insects.2. We put these two segments together and get one segment of 857bp because of the same districts of them. it had the 84%-86% homology with other insects. we conjectured it was the sequence of hsp70 mRNA of Diamondback Moth.3. Northern Blot result showed that the hsp70 in drug-fast DMB and drug-sensitive DMB at normal temperature and 37℃were examined. The expression level in drug-sensitive DMB was higher than in drug-fast DMB, in totally, the reduced level of female DMB was higher than male DMB.