Expression and Localization of Cry1Ac22 Crystal Protein from Bacillus thuringiensis W015-1 in Yeast (Saccharomyces cerevisiae)
1. Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin, 150040, China
2. Haide Institute of Tropical Agricultural Resources (HITAR), Sanya, 572025, China
3. College of Life and Technology Science, Guangxi University, Nanning, 530004, China
* These authors contributed equally to this work
Bt Research, 2010, Vol. 1, No. 3 doi: 10.5376/bt.2010.01.0003
Received: 01 Aug., 2010 Accepted: 24 Nov., 2010 Published: 31 Dec., 2010
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Preferred citation for this article:
Liu et al., 2010, Expression and Localization of Cry1Ac22 Crystal Protein from Bacillus thuringiensis W015-1 in Yeast (Saccharomyces cerevisiae), Bt Research (online), Vol.1 No.3 (DOI: 10.5376/bt.2010.01.0003)
In order to explore the expression and sub-cellular localization of Bt Cry1Ac22 insecticidal crystal protein in eukaryotic organism, we constructed eukaryotic expression vector pYES2-Cry1Ac22-GFP by fusing Cry1Ac22 and green fluorescent protein (GFP) based on the initiative pYES2 vector. The construct was transformed into yeast (Saccharomyces cerevisiae) strain INVScl to express the fusion gene promoted by GAL1. With the induction of 20%galactose, the fusion protein of Cry1Ac22-GFP was expressed as a mass of fluorescence particles in the yeast cell. SDS-PAGE analysis showed that the fusion proteins were expressed in the yeast crude succus about 130 kDa in size. The fusion proteins can generate intense fluorecense light alone with the yeast cell membrane observed under the fluorescent microscope, which indicated that Cry1Ac22 proteins were anchored in the cell membrane. This research might provide the insights and approach for studying the function of Bt insecticidal proteins as well as for tracing the trail of transgenic proteins.
Bt (Bacillus thuringiensis); Yeast (Saccharom yces cerevisiae); Cry1Ac22; Green fluorescent protein (GFP); Eukaryotic expression; Subcellular localization