Cloning and in Vitro Expression of the Human Alpha‑Fetoprotein Gene

Gulshan Stanbekova^1,2, Leila Nadirova^1,2, Daniyar Beisenov^1,2, Nailya Polimbetova^1,2 and Bulat Iskakov^1,2

¹Aitkhozhin Institute of Molecular Biology and Biochemistry, 86, Dosmukhamedov Str., Almaty, Kazakhstan.

²Institute of Plant Biology and Biotechnology, 45, Timiryazev Str., Almaty, Kazakhstan.

Corresponding Author E-mail: beisenov.d@gmail.com

DOI : http://dx.doi.org/10.13005/bbra/2361

ABSTRACT: Alpha-fetoprotein is known as a tumor marker. Strong conjugating features of the alpha-fetoprotein and its ability for specific binding with receptors of the tumor cells offers opportunity for the targeted delivery of cytostatic and cytotoxic drugs into malignant cells. Human AFP are able to induce apoptosis and to inhibit growth of the tumor cells. Technology of the recombinant DNA is the way to simplify and cheapen the production of the human AFP. Complementary DNA encoding human alpha-fetoprotein was cloned into bacterial vectors under the control of the T7 bacteriophage promoter. Various 5'- and 3'- untranslated regions (5'- and 3'-UTRs) were used as a translational enhancers. Transcription of the recombinant constructs by the T7 RNA polymerase and subsequent in vitro translation of the resulted transcripts in the wheat germ cell-free system were performed. It was shown that the human fetal blood protein could be correctly synthesized in plant cell-free system.

KEYWORDS: AFP; translational enhancers; in vitro translation; wheat germ cell-free system

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