Transformation of Kappa (?)-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. Utut Widyastuti: Department of Biology, Faculty of Mathematics and Natural Sciences, Bogor Agricultural University, Bogor, Indonesia

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

4. Alimuddin: Department of Aquaculture, Faculty of Fisheries and Marine Sciences, Bogor 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 (?)-carrageenan content, in this study ?-carrageenase gene (?car) from Pseudoalteromonas was transformed into K. alvarezii mediated by Agrobacterium tumefaciens method. The pMSH-?car 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 ?car 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-?car plasmid, while no amplification product in nontransformated (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-?car had been generated.

Page(s): 157-163

DOI: DOI not available

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

Keywords:

Keywords are not available for this article.

References:

[1] Bevan , M., 1984.Binary Agrobacterium vectors for plant transformation,Nucleic Acids Research 12 8711 -8721

[2] Campo , V.L.,D.F.Kawano,J.D.B Silva and C.I. Ivone, 2009.Carrageenans: biological properties, chemical modifications and structural analysis,Carbohydrate Polymers 77 167 -180

[3] Cheney , D.P., 2000.Agrobacterium-mediated genetic transformation of multicellular marine algae, resultant strains and their products, -

[4] Cheney , D.P.,B.Metz,J.Stiller, 2001.Agrobacterium-mediated genetic transformation in the macroscopic red alga Porphyra yezoensis,J. Phycol 37 -

[5] Clewell , D.B.,Bacterial Conjugation,WAPI(TowerID, 1993., 101313984 -43767

[6] Daud , R.F.,U.Widyastuti,Suharsono , E.,Suryati and A.,Parenrengi, 2013.Introduction of Citrat Syntase gene to Kappaphycus alvarezii using Agrobacterium tumefaciens,J.Ris. Akuakultur 8 201 -208

[7] de la Riva , G.A.,J.Gonzalez-Cabrera,C., 1998.Ayro-pardo, Agrobacterium tumefaciens: a natural tool for plant transformation,Electronic J. Biotech 1 118 -133

[8] Doty , M.S.,,M.S.,J.F.Caddy, 1987.Use of Eucheuma,FAO Technical Paper 123 -161

[9] Doyle , J.J.,J.L.Doyle, 1987.A rapid DNA isolation procedure for small quantities of fresh leaf tissue,Phytochem/ Bull 19 11 -15

[10] Fajriah , U.,E.Suryati,A.Parenrengi,U.Widyastuti, 2014.Introduction of Metallothionein Type II gene to Kappaphycus alvarezii using Agrobacterium tumefaciens,J. Ris. Akuakultur 9 377 -385

[11] Hallmann , A., 2007.Algal transgenics and biotechnology,Trans Plant J 1 81 -89

[12] Handayani , T.,Alimuddin , U.,Widyastuti,A.Parenrengi, 2014.Binary vector construction and mediated Agrobacterium tumefaciens transformation of Lysozyme gene in seaweed Kappaphycus alvarezii,Biotropia 21 80 -90

[13] Hannum , S.,Isolation, 2012.: Institut Pertanian Bogor (,PhD thesis -

[14] Hiei , Y,T.Kubo, 1997.Transformation of rice mediated by Agrobacterium tumefaciens,Plant Mol. Biol 35 205 -218

[15] Huddy , S.M.,A.E.Meyers,V.E.Coyne, 2012.Transformation of lacZ using different promoters in the commercially important red alga, Gracilaria gracilis,African J. Biotech 11 1879 -1885

[16] Khambathy , Y.,K.Mody,B.Jha, 2007.Microbulbifer elongatus comb,Nov. Biotechnol Bioprocess Eng 12 668 -675

[17] Li , F.,S.Qin,P.Jiang,Y.Wu,W. Zhang,, 2009.The integrative expression of GUS gene driven by FCP promoter in the seaweed Laminaria japonica (Phaeophyta),J Appl Phycol 21 287 -293

[18] Liberty , M. H.,G.A.Wattimena, 2008.Construction of binary plasmids carrying Cry1Ab gene and binaryplasmid transformation by triparental mating method,Zuriat 19 130 -139

[19] Lutz , C.G., 2001.Practical Genetics for Aquaculture,Pp 218 219 -

[20] McHugh , D.J.,A, 2003.: FAO,FAO Fisheries Technical Paper 441 -

[21] Michel , G.,L.Chantalat,T.Barbeyron,B.Henrissat,B.Kloareg,O.Dideberg, 2001.The κ-carrageenase of P. carrageenovora featuresa tunnel-shaped active site: A novel insightin the evolution of clan-B glycoside hydrolases,Structure 9 513 -525

[22] Neish , I.C.,Matulessi, 2005.,SEA Plant Net Technical Monograph. Makassar (ID) 48 -

[23] Provasoli , L.,A.,A, 1968.Media and prospects for the cultivation of marine algae,Jap Soc. Plant 63 75 -

[24] Rajamuddin , M.A.L,Alimuddin , U.,Widyastuti , E. Harris, and E.,Suryati, 2016.Binary vector construction of Kappa(K)-Carrageenase gene and transformation to Agrobacterium tumefaciens as mediator for seaweed transgenic generation,J ilmu dan Teknologi Kelautan Tropis 8 345 -354

[25] Rajamuddin , M.A.L.,Alimuddin , U.,Widyastuti , and I. Faizal., 2014.Evaluation of different promoters driving the GFP reporter gene in seaweed Kappaphycus alvarezii,Indonesian Journal of Biotechnology 19 129 -135

[26] Rajamuddin , M.A.L,A.A. Jaya , and R.E.,Suryati, 2010.Callus induction of Kappaphycus alvarezii for somatic embryos production,J. Ris. Akuakultur 5 211 -219

[27] Reddy,C.R.K,G.R.K. Kumar , A.K.,Siddhanta,A.Tewari, 2003.In vitro somatic embryogenesis and regeneration of somatic embryos from pigmented callus of Kappaphycus alvarezii Doty (Rhodophyta, gigartinales),J. Phycol 39 610 -616

[28] Sambrook , J.,E.F.Fritsch,T.Maniatis, 1989.A laboratory manual, -

[29] Suharsono , U.,Y.Fujisawa,T.Kawasaki,Y.Iwasaki,H.Satoh,K.Shimamoto, 2002.The heterotrimeric G protein a subunit acts upstream of the small GTPase Rac in disease resintance of rice,Proc Natl Acad Sci 99 13307 -13312

[30] Sulistiani , E.,D.T.Soelistyowati,S.A.Yani, 2012.Callus induction and filaments regeneration from callus of cottonii seaweed Kappaphycus alvarezii (Doty) collected from Natuna Islands, Riau Islands Province, 19 103 -114

[31] .,Transformation of Kapa ..... 163 -

[32] Tayco , C.C, F.A.,Tablizo , R.S.,Regalia,A.O.Lluisma, 2013.Characterization of a κ-carrageenase-producingmarine bacterium,isolate ALA B-001. Philippine JSci 142 45 -54

[33] Walker , T.L,C. Collet , and S.,Purton, 2005.Algal transgenic in the genomic era,J Phycol 41 1077 -1093

[34] Wang , J.,P.Jiang,Y.Cui,X.Deng,F.Li,J.Liu,S.Qin, 2010.Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germ plasm improvement,Aquacult Int 18 1027 -1034

[35] Wong , K.F.,J.S.Craigie, 1979.Sulfohydrolase Activity and Carrageenan Biosynthesis in Chondrus crispus (Rhodophyceae),Plant Physiol 61 663 -666

[36] Yuwono , T., 2006.,Yogyakarta (ID) 284 -

[37] Zhou , M.,J.Ma,J.Li,H.Ye,K.Huang,X.Zhao, 2008.A κ-carrageenase from a newly isolated Pseudoalteromonas-like Bacterium, WZUC10, 13 545 -551

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