Cabral Aline Diniz, Garcia Felipe Baena, Suzuki Rodrigo Buzinaro, Góis Filho Tanil Lacerda, da Costa Renata Torres, Vasconcelos Ligia Marinho Pereira, Santos Edmar Silva, Sperança Márcia Aparecida
Center for Natural and Human Sciences, Universidade Federal do ABC, Rua Arcturus, 03 - Bloco Delta, Sala 226, Laboratório 107, 09606-070 São Bernardo do Campo, SP, Brazil.
Department of Parasitology, Marília Medical School, 17519-030 Marília, SP, Brasil.
Data Brief. 2020 Sep 2;32:106259. doi: 10.1016/j.dib.2020.106259. eCollection 2020 Oct.
The data presented here is related to negative results obtained with the recombinant expression of chitinase from four species of parasites in two expression systems, performed in order to investigate the molecular characteristics of the chitinase and its possible application in leishmaniasis diagnosis. Thus, heterologous chitinase proteins were expressed in bacteria using the prokaryotic expression vector pET28a and Mach-T1, and in (Sf9) insect cells, using the eukaryotic expression system (Thermo Fisher Scientific) to produce recombinant baculoviruses to infect Sf9. Biochemical and cellular analysis of the various recombinant forms of the chitinase produced in prokaryotic and eukaryotic expression systems were performed through SDS-PAGE and Western blotting. Chitinase produced and purified from bacteria presented low yield and formed inactive aggregates. Heterologous chitinase obtained after infection of Sf9 insect cells with all the four species recombinant baculoviruses presented high yield of insoluble proteins. Dot-blot serological tests presented inconclusive results against the recombinant sp chitinases produced in both expression systems. The experiments described in this paper can help researchers to avoid errors when choosing a recombinant expression systems to produce parasites proteins for biotechnological purposes.
本文所呈现的数据与在两种表达系统中对四种寄生虫几丁质酶进行重组表达所获得的阴性结果相关，开展这些实验是为了研究几丁质酶的分子特性及其在利什曼病诊断中的潜在应用。因此，利用原核表达载体pET28a和Mach-T1在细菌中表达异源几丁质酶蛋白，并利用真核表达系统（赛默飞世尔科技公司）在（Sf9）昆虫细胞中产生重组杆状病毒以感染Sf9。通过SDS-PAGE和蛋白质免疫印迹法对在原核和真核表达系统中产生的几丁质酶的各种重组形式进行生化和细胞分析。从细菌中产生并纯化的几丁质酶产量低，且形成无活性的聚集体。用所有四种重组杆状病毒感染Sf9昆虫细胞后获得的异源几丁质酶产生了高产量的不溶性蛋白质。斑点印迹血清学试验对在两种表达系统中产生的重组几丁质酶的检测结果不明确。本文所述的实验有助于研究人员在选择用于生物技术目的生产寄生虫蛋白的重组表达系统时避免出现错误。
