Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, UPM-Serdang, Selangor Darul Ehsan, Malaysia.
Mol Biol Rep. 2011 Jan;38(1):177-82. doi: 10.1007/s11033-010-0092-4. Epub 2010 Mar 31.
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.
绿藻弯角石莼 Corda 是一种生长迅速的藻类，能够产生相当数量的类胡萝卜素和多不饱和脂肪酸。据我们所知，这是首次构建 cDNA 文库并对该物种的 ESTs 进行初步分析的报告。初级和扩增 cDNA 文库的滴度分别为 1.1×10(6)和 6.0×10(9) pfu/ml。初级文库中重组体的百分比为 97%，随机选择的 415 个原始 cDNA 克隆中有 337 个含有 600 至 1500bp 的插入片段。总共分析了 201 个大小在 390 至 1038bp 之间的个体 EST，BLASTX 评分显示，35.8%的序列被归类为强匹配，38.3%为名义匹配，25.9%为弱匹配。在具有已知假定功能的 EST 中，21.4%与基因表达有关，14.4%与光合作用有关，10.9%与代谢有关，5.5%与杂项有关，2.0%与应激反应有关，其余 45.8%被归类为新基因。本文描述的 EST 分析可以有效地从弯角石莼中分离和鉴定新基因，因此获得的序列为绿藻的广泛序列数据库做出了重要贡献。
