title: Genetic manipulation, a feasible tool to enhance unique characteristic of Chlorella vulgaris as a feedstock for biodiesel production.
abstract: Developing a reliable technique to transform unicellular green algae, Chlorella vulgaris, could boost potentials of using microalgae feedstock in variety of applications such as biodiesel production. Volumetric lipid productivity (VLP) is a suitable variable for evaluating potential of algal species. In the present study, the highest VLP level was recorded for C. vulgaris (79.08 mg l(-1 )day(-1)) followed by 3 other strains studied; C. emersonii, C. protothecoides, and C. salina by 54.41, 45 and 18.22 mg l(-1)day(-1), respectively. Having considered the high productivity of C. vulgaris, it was selected for the preliminary transformation experiment through micro-particle bombardment. Plasmid pBI 121, bearing the reporter gene under the control of CaMV 35S promoter and the kanamycin marker gene, was used in cells bombardment. Primary selection was done on a medium supplemented by 50 mg l(-1) kanamycin. After several passages, the survived cells were PCR-tested to confirm the stability of transformation and then were found to exhibit β-glucuronidase (GUS) activity in comparison with the control cells. Southern hybridization of npt II probe with genomic DNA revealed stable integration of the cassette in three different positions in the genome. The whole process was successfully implemented as a pre-step to transform the algal cells by genes involved in lipid production pathway which will be carried out in our future studies.
authors: Biotechnology and Plant Breeding Department, College of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.
authors: Ahmad Farhad
authors: AF
authors: Talebi
authors: Masoud
authors: M
authors: Tohidfar
authors: Meisam
authors: M
authors: Tabatabaei
authors: Abdolreza
authors: A
authors: Bagheri
authors: Motahhareh
authors: M
authors: Mohsenpor
authors: Seyed Kaveh
authors: SK
authors: Mohtashami
mesh_terms: D056804:Biofuels; D001711:Biotransformation; D048408:Chlorella vulgaris / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D004789:Enzyme Activation; D015870:Gene Expression; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D007612:Kanamycin / Q000494:pharmacology; D050356:Lipid Metabolism; D014170:Transformation, Genetic
chemical_list: D056804:Biofuels; D007612:Kanamycin; D005966:Glucuronidase
references: Science. 1988 Jun 10;240(4858):1534-8
references: 2897716
references: Curr Microbiol. 1999 Jun;38(6):335-41
references: 10341074
references: Bioeng Bugs. 2011 Jan-Feb;2(1):50-4
references: 21636988
references: Cell Motil Cytoskeleton. 2001 Jul;49(3):146-53
references: 11668583
references: Biotechnol Lett. 2009 Jul;31(7):1043-9
references: 19322523
references: Biotechnol Bioeng. 2009 Jan 1;102(1):100-12
references: 18683258
references: Appl Biochem Biotechnol. 2010 Mar;160(6):1674-84
references: 19424668
references: Eukaryot Cell. 2010 Apr;9(4):486-501
references: 20139239
references: Biotechnol Lett. 2010 Oct;32(10):1373-83
references: 20556634
references: EMBO J. 1987 Dec 20;6(13):3901-7
references: 3327686
references: Trends Biotechnol. 2004 Jan;22(1):45-52
references: 14690622
references: Genetics. 1998 Apr;148(4):1821-8
references: 9560396
references: Can J Biochem Physiol. 1959 Aug;37(8):911-7
references: 13671378
references: Enzyme Microb Technol. 2000 Aug 1;27(3-5):312-318
references: 10899558
references: BMC Biotechnol. 2011 Jan 21;11:7
references: 21255402
references: Mar Biotechnol (NY). 2002 Jan;4(1):63-73
references: 14961289
references: Eukaryot Cell. 2010 Aug;9(8):1251-61
references: 20562225
references: Curr Opin Biotechnol. 2010 Jun;21(3):238-43
references: 20399635