Partial characterization of Plasmodium falciparum protein kinase ABCK2 (PfABCK2)
Poster May 09, 2018
Malaria is a major threat to the public health worldwide as it is effecting populations in tropical and subtropical areas globally. Among those populations are around 40% of pregnant women and children who are susceptible to this disease. Plasmodium falciparum is the agent that causes malaria in human. There are approximately 100 protein kinases in this parasite that are involved in phosphorylation of asexual blood stage. Hence, the phosphorylation plays an important part in the development of different stages of malarial parasites.
Due to their significance in the parasite lifecycle, one of the protein kinase of P. falciparum belongs to the ABC-1 family of proteins. The bioinformatic analysis and preliminary results of PfABCK2 showed the heterologous expression of this protein. Hence, the gene of PfABCk2 was ligated into pET21a+ vector with His-tag at C-terminus and transformed into BL-21 (DE3) competent cells that were verified through Miniprep and DNA sequencing. Furthermore, this gene construct is utilized to heterologous express this protein with IPTG and afterwards purified the recombinant protein kinase using nickel affinity chromatography as shown on 10% SDS-PAGE with the expected 36 kDa protein band. Therefore, the aim of this study is to partially characterize PfABCK2 protein kinase utilizing molecular cloning, heterologous express, protein kinase activity assay and targeted screening of protein kinase inhibitors.
The novel ‘K’ DNA extraction protocol offers a reliable alternative for preparing fecal specimens for 16S rRNA gene amplicon sequencing that maintains representation of microbial populations in a sample, with the added benefits that the K method reduces hands-on time by up to 20 x for 96 sample preparations.READ MORE