RESUMO
The 2A region of the foot-and-mouth disease virus (FMDV) encodes a short sequence that mediates self-processing by a novel translational effect. Translation elongation arrest leads to release of the nascent polypeptide and re-initiation at the next in-frame codon. In this way discrete translation products are derived from a single open reading frame. Active 2A-like sequences have been found in (many) other viruses and trypanosome non-LTR retrotransposons. Exponential growth of 2A technology within the last decade has lead to many biotechnological/biomedical applications including the generation of transgenic plants/animals and genetic manipulation of human embryonic stem cells (hESCs).
Assuntos
Vírus da Febre Aftosa/química , Proteínas do Movimento Viral em Plantas/biossíntese , Proteínas do Movimento Viral em Plantas/genética , Proteínas Recombinantes/biossíntese , Proteínas Virais/química , Animais , Biotecnologia/métodos , Vírus da Febre Aftosa/genética , Engenharia Genética , Humanos , Modelos Animais , Planticorpos/metabolismo , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/genética , Biossíntese de ProteínasRESUMO
Movement proteins (MPs) that facilitate virus movement in the plants were identified in a number of plant viruses. In this study, full-length MP gene of the Chinese isolate Barley yellow dwarf virus-GAV (BYDV-GAV) was cloned and expressed in Escherichia coli. About 32% of the expressed MP was soluble providing the concentration of isopropyl-beta-D-galactopyranoside (IPTG), time of the induction, temperature and shaking speed were optimized. The soluble MP was purified using nickel-affinity column. Immune serum prepared against purified MP was used for the detection of MP in the BYDV-GAV infected leaves of oat and in the leaves of transgenic wheat plants expressing the full-length and truncated MP gene.
