Judul: Fusi Gen Kitinase Aeromonas caviae WS7b dengan Promotor sigB dari Bacillus subtilis 168 dan Ekspresinya pada Escherichia coli
Abstrak: Hama dan penyakit adalah salah satu masalah yang dihadapi oleh petani dalam sistem pertanian. Banyak metode pengendalian yang dilakukan oleh petani untuk mengurangi kehilangan hasil yang disebabkan oleh hama dan penyakit, salah satunya adalah penggunaan agens antagonis. Di alam, banyak organisme yang berpotensi menjadi agens antagonis bagi patogen tanaman. Organsime yang memiliki potensi tersebut dan digunakan dalam penelitian ini adalah Aeromonas caviae WS7b. Bakteri ini mampu mengendalikan beberapa cendawan patogen tanaman. Bakteri ini memiliki gen yang mampu menghasilkan enzim kitinase, yaitu gen chiA. Enzim ini mendegradasi dinding sel cendawan yang tersusun dari kitin. Tetapi bakteri ini tidak dapat langsung dilepas di alam sebagai agens antagonis karena bakteri ini bersifat patogen pada manusia. Sehingga perlu dilakukan rekayasa genetik terhadap gen chiA tersebut. Gen chiA ini akan digabung dengan promotor gen sigB yang terdapat pada Bacillus subtilis 168, gen sigB ini akan terekspresi sebagai respon terhadap cekaman lingkungan secara umum. Sehingga gen chiA ini akan terekspresi saat bakteri berada dalam cekaman lingkungan. Tahap awal penelitian ini adalah ekstraksi DNA total dari Bacillus subtilis 168 dan Aeromonas caviae WS7b dengan metode yang berbeda untuk masing-masing bakteri. Kemudian dilakukan amplifikasi terhadap gen chiA dan promotor gen sigB menggunakan metode polymerase chain reaction (PCR). Fragmen promotor sigB dan chiA berhasil diamplifikasi dengan PCR. Tahap berikutnya adalah penyiapan plasmid pDL2 yang akan digunakan sebagai pembawa gen rekombinan. Plasmid pDL2 diperbanyak dengan cara mentransformasikannya ke dalam bakteri Escherichia coli DH5 . Kemudian bakteri E. coli tersebut diperbanyak pada media nutrient agar (NA) yang mengandung ampisilin dengan konsentrasi 50 μg/ml. E. coli DH5 yang telah tertransformasi oleh pDL2 dibiakan pada media ampisilin dengan konsentrasi yang sama dengan saat melakukan trasnformasi. Plasmid yang telah diperbanyak dalam sel bakteri E. coli DH5 , kemudian diekstraksi. Hasil amplifikasi fragmen promoter sigB dan chiA, dipotong menggunakan enzim restriksi. Promoter gen sigB dipotong menggunakan enzim restriksi HpaI dan NheI, sedangkan gen chiA dipotong menggunakan NheI dan ApaI. Promotor gen sigB dan chiA digabungkan dengan enzim ligasi. Kedua fragmen tersebut telah berhasil digabungkan, sehingga menjadi gen rekombinan. Plasmid pDL2 yang telah dipurifikasi, dipotong menggunakan dua enzim restriksi, yaitu HpaI dan ApaI, sehingga membentuk ujung yang sama dengan gen rekombinan. Kemudian gen rekombinan yang telah dihasilkan sebelumnya, digabungkan dengan plasmid dan direkatkan dengan enzim ligasi. DNA rekombinan yang telah disambungkan dengan plasmid, berhasil ditransfomasikan ke dalam sel bakteri E. coli DH5 . Bakteri transforman ini menunjukkan aktivitas kitinase pada media kitin, karena di sekitar koloni bakteri transforman tersebut terbentuk zona bening. Hal ini menandakan bahwa gen kitinase A. caviae WS7b mampu terekspresi dengan baik di bawah promotor sigB dari B. subtilis 168.
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Judul: Kloning Gen Kitinase Aeromonas caviae WS7b pada Bacillus subtilis 168
Abstrak: Pests and diseases are the main problems in plant cultivation. Biological control is one of the control method of plant pathogen. Recently, chitinolytic bacteria has been used as biological control of plant pathogenic fungi widely. Biotechnology and genetic engineering have been successed determine the DNA sequences encoding chitinase (chiA) of A. caviae WS7b. This study to construct B. subtilis 168 transformant with promotor sigB-chiA fution. In this study, insertion of sigB-chiA fution to B.subtilis 168 chromosome was conducted by double crossover recombination into amyE locus. This research success to isolated plasmids pDL2 with inserts sigB-chiA from labolatory stock E. coli DH5α transformants and inserted to the amyE locus of B. subtilis 168 chromosome. B. subtilis 168 transformants capable to grow in medium containing chloramphenicol antibiotic, expressing chitinase on colloidal chitin agar, shown blue color with Gram staining to indicated that this transformant was B. subtilis 168.
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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