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- W2894318964 abstract "Penelitian ini bertujuan mengetahui apakah strategi yang digunakan dapat berhasil mengkloning dan mengekspresikan sekuen DNA target (daerah DBL3x gen var2csa) pada inang E.coli. Strategi kloning dilakukan melalui tahapan: perbanyakan sekuen DNA target melalui teknik PCR menggunakan primer yang didesain sendiri, peligasian sekuen DNA target pada vektor kloning pCR2.1-TOPO, dan memperbanyaknya pada inang E.coli TOP10. Strategi ekspresi dilakukan melalui tahapan: perbanyakan sekuen DNA target melalui teknik PCR menggunakan primer yang mengandung situs pengenalan enzim restriksi endonuklease tipe II: NcoI dan XhoI, pemotongan dengan enzim NcoI dan XhoI, peligasian pada vektor ekspresi pET-28a (+), dan pengekspresian pada inang E. coli BL21 (DE3) dengan induser isopropyl-1-thio-s-D-galactoside (IPTG) 1 mM. Strategi kloning berhasil mendapatkan sekuen daerah DBL3x gen var2csa berukuran 948 pb. Strategi ekspresi berhasil mendapatkan protein DBL3x berukuran 38 kDa. Kata kunci: kloning, ekspresi, DBL3x, var2csa, Escherichia coli ABSTRACT The objective of this study is to see are the strategies approach that used successfully clone and express DBL3x region of var2csa gene in E.coli host. The cloning strategies are amplifying DNA target through PCR technique using self design oligonucleotide primer, inserting into cloning vector pCR2.1-TOPO, copying in E.coli TOP10 host. The expression strategies are amplifying DNA target through PCR using self design primer with specific site for restriction enzyme NcoI and XhoI, digesting with NcoI and XhoI, inserting into pET-28a (+) expression vector, and expressing in E. coli BL21 (DE3) host by adding isopropyl-1-thio-s-D-galactoside (IPTG) 1 mM as a inducer. This study successfully cloned DBL3x region of var2csa gene with 948 pb in size and successfully expressed protein DBL3x with 38 kDa in size in E.coli system. Keywords: cloning, expression, DBL3x, var2csa, Escherichia coli" @default.
- W2894318964 created "2018-10-05" @default.
- W2894318964 creator A5040802056 @default.
- W2894318964 creator A5047507055 @default.
- W2894318964 date "2017-09-12" @default.
- W2894318964 modified "2023-09-26" @default.
- W2894318964 title "STRATEGI KLONING DAN EKSPRESI GEN var2csa PADA SISTEM Escherichia coli SEBAGAI LANGKAH PENDAHULUAN UNTUK EKSPRESI GEN PADA SISTEM EUKARIOT TANAMAN" @default.
- W2894318964 doi "https://doi.org/10.20884/1.agrin.2015.19.1.351" @default.
- W2894318964 hasPublicationYear "2017" @default.
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