ABSTRACT

Nannochloropsis

sp. is a green alga that is widely used in the aquaculture industry as a feed in Malaysia, but genetic engineering studies of this alga are still underexplored even though there is a growing interest in microalgae genetic engineering for various industrial purposes. This study aims to investigate the efficiency of three transformation methods normally done on microalgae, namely polyethylene glycol (PEG), electroporation, and glass beads on Malaysian indigenous Nannochloropsis sp. using two commercially available plasmids, pUC19 and pGEM-T easy vector as well as an amplicon of ampicillin resistance (AMPR) gene. In this study, out of three transformation methods tested, positive transformants of Nannochloropsis sp. were successfully obtained via electroporation method. Further verification via polymerase chain reaction (PCR) and sequencing confirmed that the electroporation method was found to be the sole successful method in producing transgenic lines of our locally isolated Nannochloropsis sp. Results from this study proved the efficiency of electroporation for delivery of transgene to this green alga which has been reported to be tedious. The described method also provides the gateway for developing Nannochloropsis sp. as a delivery system to aquatic organism due to its importance in the industry.

中文翻译：

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马来西亚微绿藻属物种的转化：构建转基因微藻作为水生生物疫苗输送系统的门户

摘要

微绿球藻

sp。是一种绿藻，在马来西亚水产养殖业中广泛用作饲料，但尽管人们对用于各种工业目的的微藻基因工程越来越感兴趣，但对该藻类的基因工程研究仍未得到充分探索。本研究旨在调查通常对微藻进行的三种转化方法，即聚乙二醇 (PEG)、电穿孔和玻璃珠对马来西亚本土微绿球藻的效率。使用两种市售质粒：pUC19 和 pGEM-T easy 载体以及氨苄青霉素抗性 (AMPR) 基因的扩增子。在本研究中，在测试的三种转化方法中，微拟球藻属阳性转化体。通过电穿孔方法成功获得。通过聚合酶链反应（PCR）和测序进一步验证证实，电穿孔方法是生产我们本地分离的微绿球藻转基因品系的唯一成功方法。这项研究的结果证明了电穿孔向这种绿藻递送转基因的效率，据报道电穿孔是乏味的。所述方法还为开发微拟球藻属物种提供了途径。由于其在行业中的重要性，作为水生生物的输送系统。