Cloning and nucleotide sequencing of three heat shock protein genes (hsp90, hsc70, and hsp19.5) from the diamondback moth, Plutella xylostella (L.) and their expression in relation to developmental stage and temperature.
Heat shock protein genes, hsp90, hsc70, and hsp19.5, were cloned and sequenced from the diamondback moth, Plutella xylostella (L.) by RT-PCR and RACE method. The cDNA sequence analysis of hsp90 and hsp19.5 revealed open reading frames (ORFs) of 2,151 and 522 bp in length, which encode proteins with calculated molecular weights of 82.4 and 19.5 kDa, respectively. Analysis of cDNA from hsc70 revealed an ORF of 1,878 bp coding a protein with a calculated molecular weight of 69.3 kDa. Furthermore, the analysis of genomic DNA from hsc70 confirmed the presence of introns while no introns were apparent in hsp90 and hsp19.5. Southern blot analysis suggested the presence of multiple copies of each gene family in the DBM genome. Detectable expression of hsp19.5 was observed at the pupal stage while expression of hsp90 and hsc70 was detected at both pupal and adult stages. At adult stage, females showed a higher expression of hsp90 and hsc70 than males. An increased expression was observed in all three genes after exposure to a high temperature in both sexes. These results suggest that in addition to a heat shock response, these HSP genes might be involved in other functions during the course of development in DBM.
Sonoda
Shoji
S
Research Institute for Bioresources, Okayama University, Kurashiki, Okayama, Japan. sonodas@rib.okayama-u.ac.jp
Ashfaq
Muhammad
M
Tsumuki
Hisaaki
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D015139:Blotting, Southern / Q000662:veterinary; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D050883:HSC70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D018841:HSP90 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D050883:HSC70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D006360:Heat-Shock Proteins