Article

Construction of cDNA library and preliminary analysis of expressed sequence tags from green microalga Ankistrodesmus convolutus Corda

Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM-Serdang, Selangor Darul Ehsan, Malaysia.
Molecular Biology Reports (Impact Factor: 2.02). 03/2010; 38(1):177-82. DOI: 10.1007/s11033-010-0092-4
Source: PubMed

ABSTRACT

Green microalga Ankistrodesmus convolutus Corda is a fast growing alga which produces appreciable amount of carotenoids and polyunsaturated fatty acids. To our knowledge, this is the first report on the construction of cDNA library and preliminary analysis of ESTs for this species. The titers of the primary and amplified cDNA libraries were 1.1×10(6) and 6.0×10(9) pfu/ml respectively. The percentage of recombinants was 97% in the primary library and a total of 337 out of 415 original cDNA clones selected randomly contained inserts ranging from 600 to 1,500 bps. A total of 201 individual ESTs with sizes ranging from 390 to 1,038 bps were then analyzed and the BLASTX score revealed that 35.8% of the sequences were classified as strong match, 38.3% as nominal and 25.9% as weak match. Among the ESTs with known putative function, 21.4% of them were found to be related to gene expression, 14.4% ESTs to photosynthesis, 10.9% ESTs to metabolism, 5.5% ESTs to miscellaneous, 2.0% to stress response, and the remaining 45.8% were classified as novel genes. Analysis of ESTs described in this paper can be an effective approach to isolate and characterize new genes from A. convolutus and thus the sequences obtained represented a significant contribution to the extensive database of sequences from green microalgae.

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    • "-PCR -long distance-polymerase chain reaction; RT-qPCR reverse transcription quantitative PCR. Acknowledgements : We thank Prof . Swapan Kumar Ghosh from Uttar Banga Krishi Viswavidyalaya for encouraging us in pursuing the work , and the Director , the Tocklai Experimental effective way for discovering functional genes ( Yamamoto and Sasaki 1997 , Thanh et al . 2011 ) . Identi - fication of new genes , alleles , and functional markers may help in tea improvement program using molecular breeding techniques . Although conventional breeding and propagation techniques contributed significantly to the improvement of tea for the last several decades , yet due to limitations of conventional breeding and l"

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    • "Expressed sequence tag (EST) projects provide a very useful and quick means of accessing gene sequence and expression information (Manickavelu et al., 2012). Some reports have proven that projects based on ESTs are powerful tools for both the analysis of gene expression patterns in a given tissue or at specific developmental stages (Chen et al., 2012; Tran et al., 2011; Wang et al., 2011b; Yang et al., 2009b; Zhang et al., 2011a). Moreover, ESTs from complementary DNA (cDNA) clones are inexpensive and efficient gene discovery tools (Wang et al., 2011a; Xiao et al., 2011; Yamagishi et al., 2011). "

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    • "were considered as significant (Thanh et al., 2011). Protein alignments analysis was done using the tools of Expasy, http://www.expasy.ch). "
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