Judul: Study on Over Expression of Gene Encoding Growth Hormone by Sperm Electroporation to Produce Transgenic Fast-Growing Stripped Catfish. Under direction
Abstrak: Stripped catfish (Pangasionodon hypophthalmus) is one of freshwater fish species that has high economic value in Indonesia. Limited attention has been paid to increase the growth of stripped catfish towards enhancement of its aquaculture productivity. In this research, we developed transgenesis technology to produce stripped catfish transgenic F0 as a material to obtain stable line transgenic fish with an improved growth rate character. Research was conducted in some steps: cloning and characterization of gene encoding stripped catfish growth hormone (PhGH), making of all–fish gene construction which consist of PhGH cDNA (complementary DNA) directed by carp β-actin promoter (pCcBA), developing electroporation technique using sperm mediated gene transfer (SMGT) and screening stripped catfish juvenile which carrying PhGH exogenous gene by PCR and RT-PCR methods. Complementary DNA encoding PhGH was isolated from pituitary by RT-PCR method. The size of PhGH cDNA was 1148 bp which consisted of 603 bp of ORF (open reading frame) and encoded 200 amino acid residues. PhGH gene has common characteristic like others teleost GH gene. To make all-fish gene construction of pCcBA-PhGH, PhGH cDNA was ligated with carp β-actin promoter (pCcBA). The size of pCcBA-PhGH construction was 6.6 kb. Increasing in electric field strength on sperm electroporation caused a decrease in sperm motility and embryos hatchling. Higher sperm motility and embryos hatchling were obtained at the electric field strength of 125 V/cm. Increasing concentrations of plasmid DNA caused an increase in the number of individuals carrying the exogenous PhGH gene. Electroporation using plasmid DNA concentration of 90 μg/ml effectively produced transgenic F0. Further, ectopic expression of exogenous PhGH in transgenic fish F0 at 2 months old increased 19-22.6% of their body weight compared to non-transgenic. Average body weight of 4 months juvenile treated with 90 μg/ml of DNA plasmid was 53.38% heavier than control. Therefore, stripped catfish growth rate could be increased by over expression of gene encoding growth hormone. Then, followed by stable transgenic stripped catfish generation, their culture production suggested to be increased significantly.
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Judul: Introduction and expression of tilapia (Oreochromis niloticus) growth hormone gene in catfish (Clarias sp)
Abstrak: The gene transfer technology applied in commercially important aquatic animals is to enhance genetic quality of aquaculture broodstock. This study was conducted to introduce gene encoding growth hormone (GH) in catfish embryos to improve its growth rate. In fish, microinjection was the earliest technique developed to introduce foreign DNA into fertilized eggs. However, the opaquness, stickiness and buoyancy of the embryos, the invisibility of the pronuclei, the togness of the chorion, and the higher mortality of injected eggs make this technique time consuming and requires sophisticated skills. Electropration method is able to produce mass fish transgenic. In this study, we transferred a plasmid containing GH gene of Nile tilapia (tiGH), driven by medaka β-actin promoter (mBP) into catfish using microinjection and electroporation methods, to obtain growth enhanced transgenic fish. The DNA solution (mBP-tiGH) used was 50 μg/ml in sterile distillated water. The parameter observed was survival rate of embryos (SRe), hatching rate (HR), and the percentage of individual carrying mBP-tiGH. Transgenic individual carrying tiGH was identified by PCR ( Polymerase Chain Reaction) method with specific primer for tiGH gene. The analysis of gene expression was conducted by RT-PCR. The results of research from 100 catfish embryos showed that control uninjected treatment was higher SRe and HR of eggs fertilized while the Sre and HR in electroporated-sperm was similar with control (SRe 98.5%; HR 91.2%). Percentage of catfish carrying tiGH gene by microinjection methods was 42.86% (12/28) while by electroporation methods was 87% and 93%. Germ line transmission of the transgene at first generation was 4.0 % - 8.33%. The growth of catfish in founder generation was not different between transgenic and nontransgenic. The growth of transgenic catfish at first generation were up to 7 fold higher compared with nontransgenic fish.
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Judul: Penurunan nilai albedo dan suhu permukaan dari data Terra MODIS L1B untuk klasifikasi awan
Abstrak: Clouds affect the energy balance in atmosphere through absorbtion, reflection and solar radiation process. These processes affect albedo values and the cloud surface temperature. The differences between albedo values and surface temperature can be the indicator in cloud classification and can be the separation between the cloud covered area and non-cloud covered area. Terra MODIS L1B data can be used for classifying between cloud and non-cloud using reflectant information at wavelength 0.62 μm to 0.67 μm, 0.459 μm to 0.479 μm, and 0.545 μm to 0.565 μm and the extraction of surface temperature using wavelenght 10.780 μm to 11.280 μm dan 11.770 μm to 12.270 μm. Yet, before the classification process, it is needed to look at the response of each wave lenght to the same object at the same location. Cloud‟s surface temperature can be derived from canal 31 and canal 32. The error values of both canal are 1.7 K on Januari 1st 2008, 3.3 K on March 26st 2008, 1.7 K on November 2st 2008, 5.6 K on April 6st 2008, and 12.8 K on September 24st 2008. Albedo value can be derived from band 1, band 3, band 4 since the values distribution pattern of all these canal are relatively in common. The results of the relation between reflectance bands and thermal bands show that the albedo approach method and surface temperature approach method can be used for clouds classification. The classification using albedo approach method is the based on the thickness of the clouds, while the surface temperature approach method is based on the height of clouds.
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