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Kim, Jongrae,Liu, Linpo,Hu, Zanmin,Jin, EonSeon MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.7
<P><I>Chlorella</I> has great potential as a bio-factory for production of value-added compounds. To produce the desired chemicals more efficiently in <I>Chlorella</I>, genetic tools for modification of <I>Chlorella</I> need to be developed, especially an endogenous promoter. In this study, the promoter of photosystem I protein D (<I>psaD</I>) from <I>Chlorella vulgaris</I> UTEX395 was identified. Computational analysis revealed the presence of several putative cis-acting elements, including a potential core element, and light-responsive or stress-responsive elements. Gene expression analysis in heterologous expression system in <I>Chlamydomonas</I><I>reinhardtii</I> and <I>Nicotiana</I><I>benthamiana</I> showed that <I>CvpsaD</I> promoter can be used to drive the expression of genes. Functional analysis of this promoter suggested that the initiator element (Inr) is important for its function (i.e., TATA-less promoter) and that an additional factor (e.g., downstream of the transcriptional start site) might be needed for light response. We have shown that the <I>CvpsaD</I> promoter is functional, but not sufficiently strong, both in microalgae and higher plant.</P>
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Improved plastid transformation efficiency in Scoparia dulcis L.
Kota, Srinivas,Hao, Qiang,Narra, Muralikrishna,Anumula, Vaishnavi,Rao, A.V,Hu, Zanmin,Abbagani, Sadanandam The Korean Society of Plant Biotechnology 2019 식물생명공학회지 Vol.46 No.4
The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum-specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the transformation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.
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Improved plastid transformation efficiency in Scoparia dulcis L.
Sadanandam Abbagani,Srinivas Kota,Qiang Hao,Muralikrishna Narra,Vaishnavi Anumula,A.V Rao,Zanmin Hu 한국식물생명공학회 2019 JOURNAL OF PLANT BIOTECHNOLOGY Vol.46 No.4
The high expression level of industrial and metabolically important proteins in plants can be achieved by plastid transformation. The CaIA vector, a Capsicum- specific vector harboring aadA (spectinomycin resistance), is a selectable marker controlled by the PsbA promoter, and the terminator is flanked by the trnA and trnI regions of the inverted repeat (IR) region of the plastid. The CaIA vector can introduce foreign genes into the IR region of the plastid genome. The biolistic method was used for chloroplast transformation in Scoparia dulcis with leaf explants followed by antibiotic selection on regeneration medium. Transplastomes were successfully screened, and the trans-formation efficiency of 3 transgenic lines from 25 bombarded leaf explants was determined. Transplastomic lines were evaluated by PCR and Southern blotting for the confirmation of aadA insertion and its integration into the chloroplast genome. Seeds collected from transplastomes were analyzed on spectinomycin medium with wild types to determine genetic stability. The increased chloroplast transformation efficiency (3 transplastomic lines from 25 bombarded explants) would be useful for expressing therapeutically and industrially important genes in Scoparia dulcis L.
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