Transformation of kappa (k) carrageenase gene from pseudoalteromonas in seaweed kappaphycus alvarezii [DOTY]. | [Pakistan Journal of Biotechnology • 2016]

Author(s):

1. Muh Alias L. Rajamuddin: Bogor Agricultural University, Bogor, Polytechnic Agricultural of Pangkep, South Sulawesi, Indonesia

2. Alimuddin : Department of Agriculture, Faculty of Fisheries and Marine Sciences, and Agricultural University, Bogor, Indonesia

3. Utut Widyastuti: Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Bogor, Indonesia

4. Enang Harris: Department of Aquaculture, Faculty of Fisheries and Marine Sciences, and Agricultural University, Bogor, Indonesia

Abstract:

Foreign gene transformation techniques have been applied to improve the valuable traits of farmed organism. As the first step towards increasing of kappa (x)-carrageenan content, in this study ic-carrageenase gene (war) from Pseudoalteromonas was transformed into K alvarezii mediated by Agrobacterium tumefaciens method. The pMSH-Kcar binary plasmid construct was controlled by 35S CaMV promoter and Nos terminator. DNA analysis of transformants was conducted by polymerase chain reaction (PCR) method using two sets of primer. The result showed that 36% transformed explants were sprouted and 100% of them were having the Kcar gene. The size of amplified DNA fragment from K alvarezii transformants using 35S-F and tNos-R set primers was 2,000 bp, the same size as PCR product of the positive control of pMSH-Kcar plasmid, while no amplification product in non-transformated (negative control) was found. Furthermore, PCR analysis using 35S-F and 35S-R set primers showed amplification product of 300 bp, the same size as 35S CaMV promoter sequence. Thus, K alvarezii transformants carrying pMSH-Kcar had been generated.

Page(s): 157-163

DOI: DOI not available

Published: Journal: Pakistan Journal of Biotechnology, Volume: 13, Issue: 3, Year: 2016

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