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: 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: Kerusakan bantalan rel kereta api di wilaya inspeksi 2 jalan dan bangunan perusahaan jawatan kereta api (pjka),cirebon
Abstrak: Kereta api merupakan salah satu sarana angkutan yang murah, cepat dan aman serta dapat mengangkut barang dalam jumlah besar. Kelancaran operasi sarana angkutan tersebut banyak ditentukan oleh kondisi jalan rel, termasuk kondisi bantalannya. Dalam hal ini bantalan rel kereta api dapat dibuat dari kayu, logam, atau beton. Namun demikian kayu merupakan bahan yang paling banyak digunakan sebagai pembuat bantalan rel kereta api di berbagai negara. Kenyataan menunjukkan bahwa kayu yang digunakan sebagai bantalan rel kereta api sering mengalami berbagai macam kerusakan yang menyebabkan bantalan tidak berfungsi lagi atau menurun fungsinya. Di Indonesia, informasi mengenai jenis dan tingkat kerusakan bantalan kayu masih sangat kurang, pada- hal informasi tersebut sangat penting sebagai dasar pertimbangan dalam rangka pemeliharaan jalan kereta api, agar umur pakainya (service life) lebih lama.
Keyword: bantalan rel