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Skrining Gen Cry Baru yang Aktif terhadap Nematoda dari Isolat Bacillus thuringiensis Thailand ( Screening of Novel Cry Genes Active Against Nematodes from Bacillus thuringiensis Thai Isolates)

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ABSTRAK

Ratu Elza Meirizka 24020116120008. Skrining Gen Cry Baru yang Aktif terhadap Nematoda dari Isolat Bacillus thuringiensis Thailand. Dibimbing oleh Anto Budiharjo and Sri Pujiyanto
Kristal protein yang dihasilkan oleh Bacillus thuringiensis (Bt) telah berhasil digunakan sebagai biopestisida dan tanaman transgenik di seluruh dunia. Namun, untuk melawan patogen pertanian yang paling berbahaya, yaitu nematoda puru akar, masih sangat terbatas. Kristal protein bersifat nematisidal yang diproduksi oleh Bt dapat digunakan untuk pengendalian hama yang efisien. Protein Cry 5, Cry 6, Cry 21, dan Cry 55 diketahui memiliki aktivitas nematisidal. Pada penelitian ini, 80 Bt isolat dari berbagai lokasi di Thailand diuji untuk kandungan gen cry yang memiliki aktivitas nematisidal. Skrining dilakukan dengan Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), Polymerase Chain Reaction (PCR), diligasi ke vektor kloning PJET 1.2, ditransformasi ke bakteri Escherichia coli DH5ɑ, dan digesti menggunakan enzim restriksi. Dari 80 isolat Bt Thailand yang diuji, 24 isolat (30%) memproduksi protein ±54-kDa yang setara dengan berat molekul protein Cry 6 dan 5 isolat (6%) memproduksi protein ±45-kDa yang setara dengan berat molekul protein Cry 55. Hanya satu isolat Bt (Bt 32) yang diduga positif terdapat gen cry 6.

Kata Kunci: Bacillus thuringiensis, Gen Cry Nematisidal, SDS-PAGE, PCR, Digesti.


ABSTRACT

Ratu Elza Meirizka 24020116120008.Screening of Novel Cry Genes Active Against Nematodes from Bacillus thuringiensis Thai Isolates. Supervised by Anto Budiharjo and Sri Pujiyanto
Cry proteins of Bacillus thuringiensis have been successfully used as biopesticides and transgenic crops throughout the world. However, biopesticide against the most serious agricultural pathogens, plant root-knot nematodes, are limited. Nematicidal Cry proteins produced by Bacillus thuringiensis (Bt) have been used for effiecient pest control. Cry 5, Cry 6, Cry 21, and Cry 55 were known to have nematicidal activity. In the present study, 80 Bt isolates from diverse locations in Thailand have been investigated for their nemeticidal cry gene content. Bt isolates were screened through Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), Polymerase Chain Reaction (PCR), ligated into the cloning vector PJET 1.2/blunt, transformed into Escherichia coli DH5ɑ, and diagnostic restriction enzyme digest for the presence of nematode-active cry genes. Of 80 Bt Thai isolates, 24 isolates (30%) have 54-kDa protein which was similar with the molecular weight of Cry 6 protein and 5 isolates (6%) have ±45 kDa protein which was similar with the molecular weight of Cry 55protein. Only one Bt isolate (Bt 32) was suspected to be positive for the presence of cry 6 gene.

Keywords: Bacillus thuringiensis, Nematicidal Cry Genes, SDS-PAGE, PCR, Transformation, Diagnostic Restriction Digest.

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Ketersediaan

1445B20IV

1445 B 20-iv

Perpustakaan FSM Undip (Referensi)

Tersedia

Informasi Detail

Judul Seri

BIOLOGI

No. Panggil

1445 B 20-iv

Penerbit

: ., 2020

Deskripsi Fisik

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Bahasa

Indonesia

ISBN/ISSN

-

Klasifikasi

NONE

Tipe Isi

-

Tipe Media

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Tipe Pembawa

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Edisi

-

Subjek

-

Info Detail Spesifik

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Pernyataan Tanggungjawab

Ratu Elza Meirizka

Versi lain/terkait

Lampiran Berkas

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