Obtaining of the recombinant Rhil7-Cbd fusion protein

Autor:	M. O. Usenko, M. V. Koval’chuk, D. M. Irodov, O. B. Gorbatiuk, O. V. Okunev, V. A. Kordium
Rok vydání:	2020
Předmět:	0301 basic medicine Chemistry digestive system Fusion protein digestive system diseases law.invention 03 medical and health sciences surgical procedures operative 030104 developmental biology 0302 clinical medicine Biochemistry law 030220 oncology & carcinogenesis Recombinant DNA
Zdroj:	Faktori eksperimental'noi evolucii organizmiv. 26:282-286
ISSN:	2415-3826 2219-3782
Popis:	Aim. The purpose was to obtain the recombinant fusion protein based on the human interleukin-7 (rhIL7) and cellulose binding domain (CBD). Methods. The DNA sequences encoding rhIL-7 and CBD were subcloned into the pET24a(+). Vector containing the 6His-tag sequence for further chromatographic purification of the target protein. The cells of E. coli strain BL21(DE3) were transformed with pET24-rhIL7-CBD plasmid vector. Protein synthesis was induced in clones by IPTG and by autoinduction. Results. An expression cassette of rhIL7-CBD protein has been designed. Producers of rhIL7-CBD protein were obtained. It was shown that the autoinduction protocol provides protein synthesis in E. coli (but IPTG induction doesn’t). Protein was obtained in the cytoplasmic fraction in form of inclusion bodies. In vitro method of purification of rhIL7-CBD from inclusion bodies has been worked out. Conclusions. The obtained rhIL7-CBD protein can be used for the microcrystalline cellulose immunosorbent construction for the purification of the specific polyclonal IL-7 antibodies and also for qualitative and quantitative analysis of IL-7 receptors. Keywords: IL-7, CBD, inclusion bodies, renaturation.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::e79d173a2317b75a305e51f179439dee https://doi.org/10.7124/feeo.v26.1280 Zobrazit plný text záznamu