Transformasi Gen Immunitas Gassericin A Pada

DEWAN REDAKSI
Ketua
Prof. Dr. Ir. Syamsulbahri, MS
Anggota
Prof. Dr. Ir. Moch. Munir, M.S (Unibraw-2008)
Prof. Drs. Sutiman B. Sumitro, S.U. D. Sc (Unibraw-2008)
Prof. Dr. Ir. SM Sitompul. (Unibraw-2008)
Prof. Dr. Ir. Luqman Hakim, M.S (Unibraw-2008)
Prof. Dr. Ir. Rustidja, M.S (Unibraw-2008)
Dr. Ir. Nasir Saleh (Balitkabi Deptan-2008)
Penyunting Ahli
Ir. Arifin Noor Sugiharto, M.Sc, PhD (Unibraw-2008)
Ir. Didik Suprayogo, M.Sc, PhD (Unibraw-2008)
Dr. Ir. Moch. Sahid (Balittas Deptan-2008)
Brian O Flaherty (Ahli Bhs. lnggris-2008)
Dra. Francien Herlen Tomasowa, Ph.D (Lab. Bhs. Unibraw-2008)
Drs. Adiono, M.A., PhD (Lab. Bhs. Unibraw 2008)
Penyunting Pelaksana
Drs. Sofy Permana, MSc, DSc. (Unibraw-2008)
AgustinaTri Endharti, S.Si., Ph.D (Unibraw-2008)
Sekretariat
Pujiono SH
Alamat Redaksi/Penerbit
Lembaga Penelitian Universitas Brawijaya
Jalan Veteran, Malang-65145
Telp. (0341) 575824; 551611 psw. 301; Fak. (0341) 575828
e-mail [email protected]
Transformasi Gen Immunitas Gassericin A Pada Lactobacillus gasseri Sebagai
Bukti Bahwa Vektor Ekspresi Yang Didisain Untuk Genus
Lactococcus Juga Bekerja Dengan Baik Pada Genus Lactobacillus.
Joni Kusnadi,1) Tri Ardyati,2) Purwadi3), Yasushi Kawai,4) dan Tadao Saito4)
1)
Fakultas Teknologi Pertanian Universitas Barwijaya Malang
2)
Fakultas MIPA Universitas Brawijaya Malang.
3)
Fakultas Peternakan Universitas Brawijayaya Malang
4)
Tohoku University, Tsutsumidori-Amami
yamachi, Aoba-ku, Sendai 981-8555, Japan.
Abstrak
Diterima tanggal 5 September 2005
Disetujui tanggal 27 November 2006
Plasmid pIL253-P32 merupakan plasmid yang didisain, digunakan dan bekerja dengan baik pada
bakteri asam laktat dari genus Lactococcus, misalnya pada spesies Lactococcus lactis subsp. cremoris dan
Lactococcus lactis subsp. lactis. Plasmid ini memiliki promoter kuat P32. Dengan menggunakan teknik
elektrotransformasi, pada penelitian ini kami berhasil mentransformasi plasmid pGA2 (plasmid pIL253P32 yang mengandung gen immunitas gassericin A, gaal) ke dalam bakteri Lactobacillus gasseri JCM1131.
Kami menamakan transforman yang resisten terhadap aktivitas gassericin A sebagai Lactobacillus gasseri
JYl. Konfirmasi menggunakan teknik Direct PCR dan isolasi plasmid menunjukkan bahwa fragmen DN A
yang merupakan gen immunitas gassericin A ternyata eksis dalam Lactobacillus gasseri JY1. Pembuktian
lebih lanjut melalui uji immunitas terhadap gasseicin A menunjukkan bahwa transforman Lactobacillus
gasseri JYl ternyata resisten terhadap protein antimikrobia, gassericin A. Sejauh pengetahuan kami hal ini
merupakan laporan pertama yang menunjukkan bahwa plasmid pIL253-P32 yang didisain untuk genus
Lactococcus ternyata bekerja dengan baik pada genus Lactobacillus. Penemuan ini membuka kesempatan
lebih luas bagi upaya kloning gen-gen lain pada genus Lactobacillus dengan menggunakan vektor ekspresi
yang sama.
Kata kunci: elektrotransformasi, gen immunitas gassericin A, direct PCR
Transformation of Gassericin Immunity Gene Into Lactobacillus gasseri as an Evidence that
Expression Vector for genus Lactococous also worked well in Genus Lactobacillus.
Abstract
Transformation of gassericin immunity gene into lactobacillus gasseri as an evidence that
expression vector for genus lactococous also worked well in genus lactobacillus. Plasmid pIL253-P32 is
designed, used and worked well for genus Lactococcus such as Lactococcus lactis subsp. cremoris and
Lactococcus lactis subsp. lactis. Using electrotransformation techniques, in this experiment we succesfully
transformed plasmid pGA2 (plasmid pIL253-P32 which contain gassericin A immunity gene, gaal) into
Lactobacillus gasseri JCM1131. We call the gassericin A immune transformans as Lactobacillus gasseri
JYl. Confirmation using Direct PCR and plasmid isolation showed that short fragment of gassericin
immunity gene was exist in Lactobacillus gasseri JY 1. Furthermore, gassericin immunity test also
supported that the Lactobacillus gasseri JYl transforman was resistant against proteinaceus antimicrobial,
Gassericin A. To our knowledge this is the first report that expression vector designed for Genus
Lactococcus also worked well in Genus Lactobacillus. This finding open a wide range of opportunities in
cloning of other genes into other genus of Lactobacillus using the same expression vector.
Key words : electrotransformation, gassericin A immunity gene, direct PCR
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