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: Identifikasi dan Kloning Gen CalBsyn ke dalam Plasmid pJ912-AGα-RFP di Escherichia coli
Abstrak: Sistem tampilan permukaan khamir telah menjadi teknik yang semakin popular untuk rekayasa protein dan pembuatan pustaka protein. Protein heterolog yang ditampilkan pada permukaan sel khamir diekspresikan dalam bentuk fusi dengan suatu protein penahan atau protein permukaan. Protein CALB (Candida antratica Lipase B) yang disandikan oleh gen sintetik CalBsyn dapat ditingkatkan efektivitasnya melalui ekspresi pada permukaan sel khamir. Plasmid pJ912-AGα- RFP mengandung gen penyandi protein permukaan, AGα (Agglutinin-α) dan gen penyandi protein penanda ekspresi, RFP (Red fluorescent protein). Penelitian ini bertujuan untuk memperoleh konstruksi vektor pJ912-AGα-RFP-CalBsyn untuk ekspresi protein CALB rekombinan pada permukaan sel khamir Pichia pastoris melalui kloning gen CalBsyn ke dalam plasmid pJ912-AGα-RFP pada Escherichia coli. Gen CalBsyn tanpa sinyal sekresi natifnya diperoleh dari plasmid pJ912- CALB melalui amplifikasi PCR menggunakan pasangan primer mCalB-Sal-F dan mCalB-His-Sal-R. Gen mCalBsyn yang diperoleh berukuran panjang 1004 bp. Baik gen mCalBsyn maupun plasmid pJ912-AGα-RFP dipotong dengan enzim restriksi SalI, kemudian diligasi dan ditransformasi ke dalam E. coli dengan metode kejut panas. Transformasi menghasilkan 11 koloni transforman putatif yang selanjutnya dipurifikasi menjadi koloni-koloni tunggal. Analisis PCR dengan primer AOX1-F dan mCalB-His-Sal-R menunjukkan adanya 34 koloni tunggal dari klon yang dianalisis terindikasi positif membawa plasmid rekombinan pJ912-AGα-RFPCalBsyn.
Keyword: agglutinin-α, Candida antartica Lipase B, sistem tampilan permukaan khamir, 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