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Title: Efficient transcription of the larvicidal cry4Ba gene from Bacillus thuringiensis in transgenic chloroplasts of the green algal Chlamydomonas reinhardtii
Authors: Thanate Juntadech
Kittisak Yokthongwattana
Sithichoke Tangphatsornruang
Yun-kiam Yap
Gerd Katzenmeier
Chanan Angsuthanasombat
Mahidol University. DivisionInstitute of Molecular Biosciences
Mahidol University. Faculty of Science. Department of Biochemistry and Center for Excellence in Protein Structure and Function
Keywords: Chlamydomonas reinhardtii;Chloroplast Transformation;Inverted Repeats;Bt-cry4Ba Transcript;psbA Promoter;Open Access article
Issue Date: Aug-2012
Citation: Advances in Bioscience and Biotechnology. Vol.3, 2012, 362-369
Abstract: Unicellular micro-alga Chlamydomonas reinhardtii has been recognized as a promising host for expressing recombinant proteins albeit its limited utility due to low levels of heterologous protein expression. Here, transcription of the 3.4-kb mosquito-larvicidal cry4Ba gene from Bacillus thuringiensis in transgenic C. reinhardtii chloroplasts under control of the promoter and 5’-untranslated region of photosynthetic psbA gene was accomplished. Inverted repeats in chloroplast genomes of the host strain with deleted endogenous psbA genes were selected as recombination targets. Two transformant lines were obtained by dual-phenotypic screening via exhibition of resistance to spectinomycin and restoration of photosynthetic activity. Stable and site-specific integration of intact cry4Ba and psbA genes into chloroplast genomes found in both transgenic lines implied homoplasmy of organelle populations. Achievement in cotranscription of cry4Ba and psbA transgenes revealed by RT-PCR and Northern blot analyses demonstrates the sufficiency of this system’s transcription machinery, offering the further innovation for insecticidal protein production.
Appears in Collections:MB-Article

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