Judul: Kloning Gen Proteinase Inhibitor dari Kulit Buah Kakao pada Vektor Ekspresi dengan Metode Gateway
Abstrak: Cacao (Theobroma cacao L.) is one of a high economical value estate crops as an export commodity. Eighty percent of cacao production loss was caused by Cocoa Pod Borer (CPB), Conopomorpha cramerella Snellen. Thus, cacao seed were not soldable due to its low quality. Proteinase Inhibitor (PIN) is an antinutrition substances which able to inhibit proteolytic enzyme from insect gut. A 21 kDa of cacao seed PIN gene was known of having high homology to soybean trypsin inhibitor. This research was aimed to clone PIN gene on expression vector by Gateway method. Gateway method step include design Gateway primer, amplification PIN gene with Gateway primer, recombination PIN gene on Entry and Destination vector, transformation on Escherichia coli XL- 1 Blue, confirmation PIN gene on Entry and Destination vector, and transformation PIN gene on E.coli into Agrobacterium tumefaciens. Result of research indicate that PIN gene have a measurement of 850 bp succeed inserted into expression vector with Gateway method easy and quickly. The recombinant DNA was successfully transformed into Agrobacterium tumfaciens AGL-0 strain.
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Judul: Isolasi dan Uji Ekspresi Gen Proteinase Inhibitor (PIN) dari Buah Kakao (Theobroma cacao L.).
Abstrak: Kakao (Theobroma cacao L.) merupakan salah satu tanaman perkebunan yang mempunyai nilai ekonomi tinggi sebagai komoditi ekspor. Adanya serangan hama utama kakao, yaitu Penggerek Buah Kakao (PBK), Conopomorpha cramerella Snellen menyebabkan kehilangan produksi kakao hingga 80% dan biji kakao yang terserang, mutunya menurun bahkan tidak laku dijual. Gen proteinase inhibitor (PIN) merupakan gen yang dapat menghasilkan senyawa antinutrisi yang dapat menghambat kerja enzim proteolitik di dalam perut serangga. PIN terdapat pada biji kakao dan berukuran 21 kDa dengan homologi yang cocok dengan inhibitor tripsin pada kedelai. Penelitian ini bertujuan untuk mengisolasi gen PIN dari biji kakao dan melakukan uji ekspresi untuk mengetahui tingkat ekspresi gen PIN dari jaringan kulit buah kakao yang tahan (klon Ary 2) dan tidak tahan (klon Bal 209) terhadap hama PBK. Fragmen gen PIN dari biji kakao berhasil diisolasi dengan pasangan primer spesifik gen PIN, yaitu F21 dan R11. Hasil sekuensing menunjukkan bahwa fragmen tersebut berukuran 580 bp dan analisis BLASTN membuktikan bahwa fragmen gen PIN yang berhasil diisolasi merupakan gen PIN yang mempunyai homologi dengan gen PIN penyandi protein berukuran 21 kDa pada biji kakao. Ekspresi gen PIN dari kedua klon pada buah yang memilki panjang 12 cm lebih kuat dibanding buah yang memiliki panjang 9 cm. Secara umum ekspresi gen PIN dari jaringan kulit kakao pada klon Ary lebih kuat dibanding klon Bal, dengan kata lain klon kakao yang tahan terhadap hama PBK, mengekspresikan gen PIN yang lebih kuat dibanding klon yang tidak tahan tehadap hama PBK. Berdasarkan perbedaaan ekspresi dari kedua klon, diduga gen PIN berpengaruh terhadap ketahanan buah kakao yang tahan terhadap PBK.
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Judul: Numerical modeling of the hydrodynamics of standing wave and scouring in front of impermeable breakwater with different steepnesses
Abstrak: The aim of this paper is to numerically study the effects of breakwater steepness on the hydrodynamics of standing wave and scouring process in front of impermeable breakwaters. A two-dimensional hydrodynamics model based on the Reynolds Averaged Navier–Stokes (RANS) equations and the Volume of Fluid (VOF) method was developed and then combined with an empirical sediment transport model. Comparisons with an analytical solution and experimental data showed the present model is very accurate in predicting the near bottom velocity and capable of simulating the scour/deposition patterns consistent with experimental data. It was found that the additional terms of bottom shear stress in the momentum equations are necessary to produce a physical scouring pattern. Different breakwater steepnesses produce different characteristics of standing wave, the steady streaming system, and scouring pattern in front of the breakwaters, which also affects the correlations between them. An additional analysis of the turbulence field parameters and the sediment transport rate was also performed. All these important information will be presented in details in this paper and can be worthwhile for designing the breakwater in coastal areas.
Keyword: The RANS–VOF model Standingwave Steadystreamingsystem Scouring pattern Breakwatersteepness