Identification of a Bacillus Thuringiensis Cry8Ea3 to Xin-Binding Alkaline Phosphatase from Holotrichiaparallela

摘要

In order to further study the Insecticidal Mechanism of Bt protein against Holotriciaparallela. In the study, According to the results of midgut transcriptome sequencing and RACE-PCR, the full-length hpalp gene was cloned. Sequencing analysis showed that the open reading frame of hpalp (GenBank accession NO. KY922835) was 1605 bp long, encoding 534 amino acid residues. The predicted molecular weight and isoelectric point of HpALP were 59 kDa and 5.18, respectively. Analysis of HpALP signal peptide with 21 amino acids, GPI anchor point is located in the C-terminal D~(514), and has two N-glycosylation sites: N~(100), N~(296). The results of Ligand blot showed that the binding of HpALP with Cry8Ea3 toxin. Transcriptional analysis of hpalp in different tissues of H. parallela larvae was performed by qRT-PCR, which revealed that the hpalp was primarly expressed higher in midgut, but lower in the foregut.