Judul: Delesi Sinyal Sekresi Gen CalBsyn dalam Plasmid pGAPZα-proCalBsyn-egfp melalui Teknik Mutagenesis Terarah.
Abstrak: ProCalBsyn merupakan gen penyandi Candida antartica lipase B yang dibuat secara sintetik dalam plasmid pGAPZα-proCalBsyn-egfp (4839 bp). Sekuen proCalBsyn masih mengandung sinyal sekresi native (ss-CalBsyn), sementara plasmid pGAPZα memiliki sinyal sekresi mating-factor-α.(MFα). Tujuan dari penelitian ini adalah mendelesi sinyal sekresi ss-CalBsyn, mengintroduksi situs EcoRI serta transformasi plasmid mutan ke E.coli TOP10F’. Delesi ss-CalB dilakukan dengan teknik mutagenesis terarah melalui proses PCR. Sepasang primer F-mCalB dan R-mCalB serta high-fidelity DNA-polymerase G-Hifi digunakan untuk amplifikasi. Hasil amplifikasi PCR diperoleh plasmid mutan berupa DNA linear berukuran 4780 bp. Setelah direstriksi dengan enzim EcoRI dan ligasi, plasmid mutan ditransformasikan kembali ke E.coli. Transformasi menghasilkan lebih dari 60 koloni transforman. Analisis PCR menunjukkan 3 dari 9 klon hasil isolasi plasmid yang dianalisis terindikasi membawa plasmid mutan yaitu klon 7, 19 dan 20. Ketiga klon tersebut menunjukkan adanya pita gen mCalBsyn (970 bp).
Keyword: mutagenesis terarah, mutasi delesi, sinyal sekresi native CalB, proCalBsyn, plasmid pGAPZα

Judul: Konstuksi Vektor Dan Ekspresi Protein Rekombinan Lipase Candida Antarctica (Calb) Dengan Cellulose Binding Module (Cbm) Pada Pichia Pastoris
Abstrak: Gen CalBsyn telah dikonstruksi secara sintetik untuk menyandi Candida antarctica lipase B (CALB). Enzim tersebut memiliki peranan penting sebagai biokatalis yang efektif di bidang bioteknologi dan industri. Sekuens gen CalBsyn telah dimodifikasi dengan mengintroduksi mutasi pada tiga asam amino, yaitu V210I, A281E, dan V221D untuk meningkatkan termostabilitas enzim tersebut. Fragmen gen cellulose binding module (cbm) endoglukanase I (egl1) dari Trichoderma reesei difusikan pada ujung karboksil gen CalBsyn pada plasmid rekombinan pGAPZα-CalBsyn. CBMmerupakan suatu domain protein yang umum dijumpai pada enzim selulase dan berfungsi dalam pengikatan enzim pada matrik selulosa. Dalam penelitian ini CBM digunakan untuk memfasilitasi imobilisasi CALB pada matrik selulosa melalui fusi dari dua protein tersebut. Penelitian ini bertujuan mengkonstruksi vektor ekspresi gen CalBsyn yang difusi dengan cbm dan mengekspresikan CALB pada Pichia pastoris SMD1168H. Baik plasmid maupun gen cbm dipotong dengan enzim ClaI. Plasmid rekombinan pGAPZα-CalBsyn-cbm dikonstruksi dengan cara meligasikan gen cbm ke dalam plasmid pGAPZα-CalBsyn. Plasmid rekombinan pGAPZα-CalBsyn-cbm diklon dan diseleksi pada Eschericia coli TOP10F’. Plasmid rekombinan pGPAZα-CalBsyn telah berhasil diintroduksi ke dalam genom P. pastoris SMD1168H melalui metode elektroporasi. Efisiensi transformasi yang diperoleh sebesar 1,01 x 102 cfu/μg DNA. Analisis ekspresi protein rekombinan dilakukan terhadap beberapa galur rekombinan P. pastoris terpilih. Uji aktivitas lipase secara kualitatif pada media seleksi menunjukkan bahwa galur transforman P. pastoris yang diperoleh positif mensekresikan lipase rekombinan (CALB) dan memiliki aktivitas lipolitik. Uji aktivitas lipase secara kuantitatif dari salah satu galur transforman P. pastoris menunjukkan bahwa aktivitas enzim mencapai 6,35 Units/ml pada jam ke-48. Analisis protein dengan SDS-PAGE menunjukkan bahwa CALB berhasil diekspresikan secara ekstraseluler dengan ukuran sekitar 45 kDa.
Keyword: Candida antarctica lipase B, cellulose binding module, recombinant Pichia pastoris

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