Judul: Pemurnian Dan Pengujian Aktivitas Enzim Rekombinan Dna Polimerase Pyrococcus Furiosus Asal Gen Sintetik
Abstrak: Enzim asam deoksiribonukleat (DNA) polimerase merupakan enzim yang berperan sangat luas dalam perkembangan bioteknologi dan biologi molekuler, terutama dalam aplikasi teknik reaksi rantai polimerase (PCR). Penelitian ini bertujuan memurnikan enzim rekombinan DNA polimerase Pyrococcus furiosus (Pol-pfu) agar memperoleh aktivitas yang baik ketika diaplikasikan melalui teknik PCR. Pol-pfu kasar dimurnikan menggunakan metode kromatografi afinitas ion logam. Pemurnian ini menghasilkan 1 pita spesifik pada SDS-PAGE dengan bobot molekul sekitar 92 kD. Pol-pfu dengan konsentrasi 50 μg/mL berhasil digunakan untuk mengamplifikasi sampel DNA yang memiliki ukuran pasangan basa sebesar 200 hingga 2000. Hal ini menunjukkan bahwa Pol-pfu memiliki aktivitas sesuai dengan yang diharapkan, yaitu mampu mengamplifikasi sampel DNA berukuran pasangan basa kecil hingga besar.
Keyword: DNA polimerase, kromatografi afinitas, Pyrococcus furiosus, pemurnian enzim

Judul: Development In House of Taq DNA Polymerase Recombinant Based
on Synthetic Gene and Its Applications
Abstrak: Biologi molekuler merupakan salah satu disiplin ilmu dasar yang berperan penting dalam menunjang berkembangnya bioteknologi modern. Salah satu teknik yang digunakan dalam bidang biologi molekuler adalah teknik PCR yang membutuhkan enzim termostabil tahan panas. Taq DNA polimerase adalah enzim termostabil yang banyak digunakan untuk amplifikasi DNA dalam teknik PCR. Enzim ini awalnya dikarakterisasi dan diisolasi dari bakteri termofilik yaitu Thermus aquaticus, namun terdapat kesulitan untuk mengembangkan enzim ini jika menggunakan sistem inang asli. Penelitian ini bertujuan untuk mengembangkan enzim Taq DNA polimerase rekombinan berbasis gen sintesis menggunakan sistem ekspresi E.coli. Cetakan runutan nukleotida gen Taq DNA polimerase diperoleh dari data GeneBank J04639.1 dan optimasi kodon dilakukan sebelum gen disisipkan ke dalam vektor pET151/D-TOPO. Sintesis gen dilakukan di GenArt Gene Synthesis (Thermofisher Scientific, USA). Gen sintetik Taq DNA polimerase kemudian diekspresikan menggunakan bakteri E.coli BL21 dan diinduksi menggunakan IPTG (Isopropyl-β-D thiogalactopyranoside). Pemurnian enzim rekombinan Taq DNA polimerase dilakukan menggunakan kromatografi afinitas Ni2+-NTA dan enzim murni yang dihasilkan diaplikasikan pada teknik PCR. Gen Taq DNA polimerase berukuran sebesar 2685 pb berhasil disisipkan ke dalam vektor pET151/D-TOPO dan analisis hasil ekspresi dilakukan menggunakan teknik SDS PAGE dan menunjukkan protein target yang spesifik berukuran 100.9 kDa. Konsentrasi dan aktivitas enzim yang dimurnikan berturut turut adalah 5.17 mg/mL dan 4.647 U/μL. Aplikasi enzim ini pada teknik PCR menunjukkan bahwa enzim ini dapat mengamplifikasi gen target mulai dari 200 pb hingga 3500 pb amplikon dengan konsentrasi DNA cetakan minimum 10 ng/μL. Hal ini mengindikasikan bahwa in house enzim Taq DNA polimerase rekombinan yang dibuat berdasarkan gen sintesis telah berhasil diekspresikan, dimurnikan, dan diterapkan untuk teknik PCR dan dapat diterapkan dalam bidang bioteknologi khususnya dalam teknik PCR/RT-PCR
Keyword: Codon-optimized, E.coli BL21, Ni2+-NTA purified system, Synthetic Taq DNA polymerase

Judul: Characteristic of Overshooting Top (OT) Based on  Himawari  8  Satellite  as  a  Convective  Cloud  Feature  that  Generates  Extreme Weather in the Java Island Region
Abstrak: Overshooting Top (OT) is a feature at the top of a convective cloud that is associated with extreme weather. Research on OT in tropical Indonesia is still very limited. The existence of geostationary satellites with wide area coverage and fast frequency observations is useful for observing the fast dynamics of the OT life cycle. Therefore, this study intends to analyze the relationship between OT and extreme weather, updraft and thermodynamic conditions of OT when extreme weather occurs, the best objective method for detecting OT in the day time and night time, and the spatiotemporal distribution of OT in the tropical region of Java Island during the 2018-2021 period. Based on visual analysis using visible channel and Himawari 8 sandwich products, 87% of extreme weather events in Java were accompanied by the appearance of OT. It indicates that the presence of the OT pattern is a convection cloud pattern or feature that is closely related to extreme weather events for the Java Island region. The existence of a time lag between the appearance of OT and extreme weather events on Java Island indicates that the OT of convection cloud can be used as a precursor or nowcasting for the appearance of extreme weather. Most of the convective cloud OT features during extreme weather events on Java Island have altitude that were only able to exceed the altitude of the equilibrium level and few were able to exceed the tropopause level of 9 km – 16 km. The difference in atmospheric conditions during extreme weather events on Java Island, which is in the tropics and subtropics, is a strong and unstable convective energy condition characterized by a larger CAPE, as well as unstable KI, LI, SI and TTI values. It is offset by kinematic factors in the form of a weak vertical wind shear (bulk wind shear 0-6 km) and a higher freezing point height. The difference between extreme weather events in the Java Island region and other tropical regions compared to subtropical regions is the strong updraft speed of convection clouds. The IRW Texture COMB algorithm based on the IRW-Texture Gradient modification and the COMB algorithm shows a significant increase in quality for detecting OT and non-OT pixels in Java. Rainy days with rainfall > 50 mm/day are very common in areas with an altitude of > 500 m. On the other hand, the level of convection activity which is indicated by the presence of OT and lightning is significantly reduced in this area. OT of convective clouds appear more frequently during the DJF season and less frequently during the JJA season. OT often appears during the day time and rarely at night time. Most OTs appear in the central part, away from the south and north coasts of Java Island during the day, while OTs that appear near the south and north coasts of Java Island can appear in the morning or afternoon.
Keyword: convective clouds, extreme weather, Overshooting Tops, Himawari 8 Satellite