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将马来西亚的一种 :转化为构建转基因微藻作为水生生物疫苗传递系统的门户。

Transformation of a Malaysian species of : gateway to construction of transgenic microalgae as vaccine delivery system to aquatic organisms.

机构信息

Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia , Serdang, Malaysia.

Aquatic Animal Health and Therapeutics Laboratory (Aquahealth), Institute of Bioscience, Universiti Putra Malaysia , Serdang, Malaysia.

出版信息

Bioengineered. 2020 Dec;11(1):1071-1079. doi: 10.1080/21655979.2020.1822106.

Abstract

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 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 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 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 sp. as a delivery system to aquatic organism due to its importance in the industry.

摘要

sp. 是一种绿色藻类,在马来西亚的水产养殖业中被广泛用作饲料,但即使人们对微藻基因工程在各种工业用途中的应用越来越感兴趣,对这种藻类的基因工程研究仍不够深入。本研究旨在探讨三种通常用于微藻的转化方法(聚乙二醇(PEG)、电穿孔和玻璃珠)在使用两种市售质粒(pUC19 和 pGEM-T easy 载体)和氨苄青霉素抗性(AMPR)基因扩增子的情况下,对马来西亚本土 sp. 的转化效率。在本研究中,在所测试的三种转化方法中,通过电穿孔法成功获得了 sp. 的阳性转化体。通过聚合酶链反应(PCR)和测序进一步验证,证实电穿孔法是唯一能够成功生产我们本地分离的 sp. 转基因株系的方法。本研究结果证明了电穿孔法在将转基因递送到这种已被报道为繁琐的绿色藻类中的有效性。由于 sp. 在该行业中的重要性,所描述的方法也为将其开发为水生生物的递送系统提供了途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6dc1/8293953/1d3e30a2330b/KBIE_A_1822106_F0001_OC.jpg

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