Published date: Feb 11 2020

Journal Title: KnE Life Sciences

Issue title: The 2019 International Conference on Biotechnology and Life Sciences (IC-BIOLIS)

Pages:

DOI: 10.18502/kls.v5i2.6447

Febriana Dwi WahyuniDepartment of Biotechnology, Faculty of Health, Universitas Esa Unggul

Cindy FransiscaDepartment of Biotechnology, Faculty of Health, Universitas Esa Unggul

Henny Saraswatihennysaraswati@esaunggul.ac.idDepartment of Biotechnology, Faculty of Health, Universitas Esa Unggul

Bacillus thuringiensis is a biological biopesticide that was used for defense against pests. In B. thuringiensis there is a Cry protein that is only toxic to certain insects. This Cry protein is encoded by cry gene. There are many types of cry genes that have been identified, one of which is cryIII gene that is toxic to insects from the Coleoptera group as pests in sweet potato (Ipomoea batatas). The aim of this study was to amplify the cryIII gene from local isolate of B. thuringiensis. The method that can be used  for cryIII gene amplification is Polymerase Chain Reaction (PCR). The primer pair is one of the important factors that determine the success of PCR. From a number of designed primers, the primer pair selected to be used in this study is Cry3B forward 5’-AAAGTGCGGCTATTCGACCA-3’ and Cry3B reverse 5’-CACTTCATCCTGTGACGCCT-3’. This primer pair successfully amplified cryIII gene and showed a DNA band with molecular size approximately 914 base pairs. Gradient PCR needs to be done for optimizing specific amplification of cryIII gene.

Keywords: PCR, primer, sweet potato

[1] Saraswati NPA. Deteksi dan Identifikasi Gen Cry4 pada Isolat Bacillus thuringiensis Daerah Bogor dan Sekitarnya. Inst Pertan Bogor, Bogor. 2007.


[2] Bahagiawati. Penggunaan Bacillus Thuringiensis sebagai biolarvasida. Bul AgroBio. 2003;5(1):21-28.


[3] Suryanto D. Amplifikasi gen Cry1 dan analisis genom isolat Bacillus thuringiensis lokal. J Biol Res.2017;15(1):1-4. doi:10.23869/bphjbr.15.1.20091


[4] Malik K, Muhammad M, Talpur A. Cloning and Expression of a cry III Gene Isolated from the Local Habitat into a Modified Strain of Bacillus thuringiensis. IOSR J Pharm Biol Sci. 2013;7(5):87-95.


[5] Ceron J, Ortiz A, Quintero R, Guereca L, Bravo A. Specific PCR primers directed to identify cryI and cryIII genes within a Bacillus thuringiensis strain collection. Appl Environ Microbiol. 1995;61(11):3826-3831.


[6] Suryadi PT, Ratnayani K, Yowani SC. Desain Primer untuk Amplifikasi Gen katG Multidrug Resistance Tuberculosis (MDR-TB) dengan Metode Polymerase Chain Reaction (PCR). J Kim. 2014;8(1):77-82.


[7] Septiari IGAA, Yustiantara PS, Yowani SC. Analisis Primer untuk Amplifikasi Promoter inhA Multidrug Resistance Tuberculosis (MDR-TB) dengan Metode Polymerase Chain Reaction (PCR). J Kim. 2015;9(1):117-123.


[8] Zuker M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Reasearch. 2003;31(13):3406-3415. doi:10.1093/nar/gkg595


[9] Suparman, Ahmad H, Ahmad Z. Desain Primer PCR Secara in silico untuk Amplifikasi Gen COI pada Kupu-kupu Papilio ulysses Linnaeus dari Pulau Bacan. J Pendidik Mat dan IPA. 2016;7(1):14-25.


[10] Dewi RW, Dewi VR, Yowani SC, Yustiantara PS. Desain Primer untuk Amplifikasi Regio Promoter Gen inh A Isolat P016 Multidrug Resistance Mycobacterium tuberculosis dengan Metode Polymerase Chain Reaction. J Farm Udayana. 2018;7(1):34-39.


[11] Safitri TA, Nurul D, Patty J, Saraswati H. Gen L1 HPV 16 dan 18 Sebagai Dasar Dalam Desain Primer untuk Deteksi Kanker Leher Rahim dengan in-House Multiplex PCR. Indones J Biotechnol Biodivers. 2018;2(2):67-71.