Prokaryotic expression and expression profiling of calmodulin genes in mealybug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)
[Objective] The research investigated the expression of calmodulin gene in Phenacoccus solenopsis (PsCam) at different developmental stages and its heterologous expression in vitro to improve the further exploration of its physiological function.[Method] The ORF of PsCam was cloned,the amino acid sequence was derived,and its structure was analyzed using bioinformatics methods.The prokaryotic expression vector was constructed before being transformed into E.coli for expression and purification.Its relative expression levels at different instars were analyzed using real-time PCR.[Result] The amino acid sequence indicated that there were two EF-hand domains and 13 Ca2+ binding sites but no signal peptide in the protein of PsCam.The prokaryotic expression vector of pET28a-PsCam was successfully built.The relative molecular mass of the obtained protein after induced expression was approximately 17 ku.Expression of PsCam mRNA was observed in different instars of P.solenopsis.The expression in mature females was the lowest,while the expression in one year old instar larvae was the highest,which was 7.1 times of that in mature females.[Conclusion] P.solenopsis calmodulin protein was successfully expressed and the expression levels in different instars were different.
NETL
NSTL
万方数据
维普
