Hasil Pencarian  ::  Simpan CSV :: Kembali

Abstrak :
Latar Belakang : Sistem in vitro pendeteksi interaksi protein Vif HIV-1 dengan Apobec3G akan mempermudah identifikasi obat anti HIV-1 yang dapat meghambat replikasi HIV-1 melalui fungsi protein intrinsik Apobec3G. Protein Apobec3G rekombinan yang diperoleh melalui ekspresi pada sistem prokariota dapat digunakan bersama-sama dengan protein vif rekombinan untuk pengembangan sistem identifikasi substansi penghambat interaksi Apobec3G dan Vif HIV-1 dalam rangka eksplorasi protein bahan alam sebagai penghambat infeksi HIV. Metode : Gen Apobec3G diklona ke dalam vektor plasmid pGEX6P-1 dalam E.coli TOP10, kemudian dilanjutkan dengan ekspresi pada sel E.coli BL21 untuk memperoleh protein rekombinan. Proses induksi dilakukan pada suhu 37°C dengan konsentrasi IPTG 0,5 mM, waktu induksi 2 dan 4 jam. Hasil : Gen apobec3G telah berhasil diklona ke dalam vektor ekspresi prokariot, tetapi protein belum berhasil diekspresikan. ...... Background: A system for detection of in vitro interaction of HIV-1 Vif protein with apobec3G protein will facilitate the identification of novel anti HIV-1 drug that inhibit the virus replication through functional intrinsic Apobec3G protein. Recombinant Apobec3G protein that is obtained through expression in prokaryotic expression system can be utilized in combination with recombinant HIV-1 Vif protein for the development of a sistem for identification of an inhibitory substance for interaction of Apobec3G and HIV-1 Vif, in order to explore the potential of natural substances as inhibitors of HIV infection. Methods: The gene encoding the Apobec3G protein was cloned into the plasmid vector pGEX6P-1 in Top10 E. coli, followed by expression in BL21 E. coli to obtain the recombinant protein. Induction of expression was performed at 37oC with IPTG concentration of 0.5 mM for 2 and 4 hours. Results: The gene encoding the Apobec3G has been successfully cloned in the prokariotic expression vector, however the expression of the corresponding recombinant protein has not been successful.

Abstrak :
Kura-kura rote leher ular (Chelodina mccordi) merupakan spesies endemik dari Pulau Rote, yang tergolong Critivally Endangered-Possibly Extinc in the Wild (CRPEW) berdasarkan IUCN. Rendahnya kepadatan C. mccordi menjadi tantangan dalam pemantauan secara visual, sehingga dibutuhkan pendekatan molekuler sebagai metode alternatif. Namun, primer spesifik untuk C. mccordi belum pernah dikembangkan. Penelitian dilakukan dengan mengembangkan markah genetik yang menargetkan gen ND2 mtDNA pada sampel eDNA dari air melalui metode qPCRHRM, serta menguji sensitivitas dan spesifisitas primer. Bahan uji yang digunakan adalah air kolam C. mccordi, C. expansa, campuran (C. mccordi dan C. expansa), dan air keran. Hasil perancangan primer didapat panjang produk 179 bp dengan menganalisis basa unik yang hanya diekspresikan oleh C. mccordi. Analisis kualitatif dengan PCR dan visualisasi elektroforesis, serta validasi dengan sekuensing menunjukkan primer memiliki spesifisitas yang tinggi terhadap C. mccordi. Primer yang dirancang hanya mampu mengamplifikasi hingga dilusi 3 tingkat dengan faktor dilusi 10x. Efisiensi primer tergolong baik dengan nilai 100%, dan persamaan regresi linear (y= -3,32x + 34,127), serta R2 sebesar 0,9801. Analisis HRM dilakukan untuk mendeterminasi C. mccordi berdasarkan suhu luruh (80—81˚C). Hasil pengujian sensitivitas dan spesifisitas sebesar 81,25% dan 62,5% menunjukkan bahwa primer tidak direkomendasikan untuk analisis kuantitatif dengan qPCR-HRM. Pengembangan desain primer spesifik-spesies perlu dirancang kembali dengan gen target lain seperti COI dan Cyt-b untuk pendeteksian C. mccordi melalui eDNA. Meskipun demikian, primer yang dirancang tetap bisa digunakan untuk analisis kualitatif. ......The Roti Island Snake-necked Turtle (Chelodina mccordi) is an endemic species of Roti Island, classified as Critically Endangered-Possibly Extinct in the Wild (CRPEW) according to the IUCN. The low density of C. mccordi poses a challenge for visual monitoring, necessitating a molecular approach as an alternative method. However, specific primers for C. mccordi have not been developed. The research involved the development of genetic marker targeting the ND2 mtDNA gene in eDNA samples from water using the qPCR-HRM method and testing the sensitivity and specificity of the primers. Test materials included water from C. mccordi, C. expansa, a mixture (C. mccordi and C. expansa), and tap water. The designed primer obtained a product length of 179-bp by analyzing unique bases specific to C. mccordi. Qualitative analysis with PCR and electrophoresis visualization, then validated by sequencing, showed high primer specificity for C. mccordi. The designed primers could only amplify up to a 3-level dilution with a 10x dilution factor. The qPCR reaction efficiency was considered good at 100%, with a linear regression equation (y = -3.32x + 34.127) and an R2 of 0.9801. HRM analysis was carried out to determine C. mccordi based on the melting temperature (80—81˚C). Sensitivity and specificity test results of 81.25% and 62.5% indicated that the primers are not recommended for quantitative analysis with qPCR-HRM. The redesign of species-specific primer with other target genes such as COI and Cyt-b for C. mccordi detection via eDNA is needed. Nevertheless, the designed primers can still be used for qualitative analysis.

Abstrak :
Analysis of Genetic Association Studies, textbook in statistical genetics and genetic epidemiology, and a reference book for the analysis of genetic association studies. The book is applicable to the study of statistics, biostatistics, genetics and genetic epidemiology. In addition to providing derivations, the book uses real examples and simulations to illustrate step-by-step applications. Introductory chapters on probability and genetic epidemiology terminology provide the reader with necessary background knowledge.