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: Introduksi dan Ekspresi Gen Hormon Pertumbuhan pada Ikan Mas Transgenik
Abstrak: Common carp Cyprinus carpio is one of the economically valuable aquaculture fish species in Indonesia. This research was aimed to produce transgenic common carp that grow rapidly by over-expressing the tilapia growth hormone (tiGH) gene. Trangenic common carp were produced by sperm-mediated gene transfer using electroporation method. In this study, two electroporation programs that developed by other researcher were applied to obtain a suitable electroporation method for common carp. The electroporated sperm were then used to fertilize eggs to produce F0 transgenic fish. Transgenic F0 carrying tiGH in their sperm were crossed with non-transgenic fish to produce transgenic first (F1) generation. Identification of transgenic fish was performed by PCR method using specific primer for tiGH gene. Expression of tiGH was analyzed using RT-PCR method and growth measurement at F1 fish generation. The results showed that electroporation of sperm using 50 Volt/cm, and DNA concentration of 25 ng/μl was produced higher percentarge of fish carrying tiGH in their gamet compared to that of another electroporation program (40 Volt/cm, DNA concentration of 50 ng/μl). Five sexually matured males F0 transgenic fish were selected and mated with non-transgenic to produce F1 generation. Percentage of F1 progenies carrying tiGH was 47,2-66,7%. Variated measure of F1 genereations was 10,1->12,0 cm size (22,5%-42,9%), 8,1->10,0 cm size (14,3%-42,9%), 6,1->8,0 cm size (11,9%-29,2%), 4,1->6,0 cm size (0,0%-39,3%). Increasing of growth of F1 was about 1,4-1,87 times for non transgenic fish. Survival of transgenic and non-transgenic F1 was generally similar. RT-PCR analysis showed that the F1 transgenic fish have tiGH gene expression in caudal fin, while no expression in non-transgenic fish were found. Thus, the increased body weight of F1 transgenic common carp was associated with expression of tiGH gene, and this fish can be used as broodstock for the production of fast-grow stable transgenic common carp.
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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