Pramesti, DiasIdha (2009) Identifikasi Gen Pengkode Thaumatin-Like Protein (TLP) pada Beberapa Varietas Apel (Malus sylvestris Mill.). Magister thesis, Universitas Brawijaya.
Abstract
Thaumatin-like protein (TLP) merupakan protein yang pada umumnya terakumulasi pada saat pematangan buah dan terkait dengan akumulasi gula. Gen pengkode protein TLP tersebut telah diidentifikasi pada beberapa tanaman buah, antara lain pepper dan apel varietas Prima, Fiesta serta Fuji. Akan tetapi belum terdapat informasi tentang keberadaan TLP pada tanaman apel yang ditanam di Indonesia diantaranya Kota Batu. Oleh karena itu penelitian dilakukan untuk mengetahui keberadaan gen pengkode TLP, sekuen gen pengkode serta tingkat kesamaan sekuen gen pengkode TLP pada tanaman apel varietas Anna, Manalagi, Rome Beauty, Wanglin dan Australi. Sampel tanaman apel dari setiap varietas yang berupa daun muda diperoleh dari kebun apel milik petani di Kota Batu. Isolasi DNA genom menggunakan metode Doyle and Doyle dilakukan di Laboratorium Biologi Molekuler Jurusan Biologi Fakultas Matematika dan ilmu Pengetahuan Alam Universitas Brawijaya Malang . Isolat DNA yang memiliki kemurnian cukup tinggi selanjutnya diamplifikasi dengan teknik PCR ( Polymerase Chain Reaction ) menggunakan pasangan primer Mal d 2.01B dan PepTLP. Kedua primer tersebut disusun berdasarkan sekuen gen pengkode TLP Malus domestica cultivar Prima ( Mal d 2.01B ) dan Capsicum annuum L. ( PepTLP ). DNA hasil amplifikasi tersebut kemudian dibaca urutan sekuen basa nukleotidanya dengan teknik sekuensing. Pembacaan sekuen gen pengkode TLP dilakukan di Balai Pengkajian Bioteknologi (BPPT) menggunakan mesin ABI 3130 Genetic Analyzer. Analisis sekuen gen pengkode TLP dilakukan dengan program BLAST. Hasil amplifikasi DNA genom menunjukkan bahwa keberadaan gen pengkode TLP dapat dideteksi dengan teknik PCR menggunakan primer Mal d 2.01B menghasilkan pita amplifikasi berukuran sekitar 600 bp. Primer PepTLP tidak dapat mengamplifikasi DNA genom tanaman apel, menunjukkan bahwa pada apel varietas Rome Beauty, Manalagi, Australi, Wanglin serta Anna terdapat gen pengkode TLP, Mal d 2.01B dan tidak terdapat gen PepTLP . Hasil sekuensing menunjukkan bahwa sekuen gen Mal d 2.01B pada kelima tanaman apel tersebut memiliki rata-rata tingkat kesamaan atau nilai similaritas 98% sehingga terdapat perbedaan nilai sebesar 2%. Perbedaan sekuen antar varietas apel dalam hal susunan basa nukleotida diduga disebabkan oleh perbedaan alel pada gen pengkode TLP masing-masing varietas.
English Abstract
Thaumatin-like protein (TLP) is one of highly accumulated protein during fruit ripening. expression of TLP gene has been identified having correlation with sugar accumulation too. Thaumatin-like protein gene has been identified in several fruit plant such as pepper, apple var. Prima, Fiesta and Fuji. re is no information about its existance in Indonesians cultivated apple yet. So, objectives of this research were to know existence, sequence and similarity of TLP gene on apple fruit varieties, Anna, Manalagi, Rome Beauty, Wanglin and Australi. materials of genomic DNA extraction are apple young leaves which grown in Batu. extraction of genomic DNA used Doyle and Doyle method in Molecular Biology Laboratory of Science Faculty-Brawijaya University. existence of TLP gene was detected by PCR technique using Mal d 2.01B and pepTLP primers. Both of primer pair was based TLP gene sequence of Malus domestica cultivar Prima ( Mal d 2.01B ) and Capsicum annuum L. ( PepTLP ). Sequencing to know TLP gene sequence were conducted by Balai Pengkajian Bioteknologi (BPPT) using ABI 3130 Genetic Analyzer machine. Thaumatin-like protein gene sequences were analyzed by BLAST programme. result shows that both of primer pairs, Mal d 2.01B and pepTLP have different capability to amplify all of samples. PCR using Mal d 2.01 primer resulted in 600 bp band but re is no band for PepTLP. It indicated that apple has Mal d.01B gene. Sequencing results were analyzed by BLAST programme showed that all of apple TLP gene have similarity sequence. similarity were about 98%. Although Mal d 2.01B of apple fruit varieties, Anna, Manalagi, Rome Beauty, Wanglin and Australi highly similar, re are some different bases caused 2% different value. It seems that sequence differences may be caused by different TLP gene allele of each variety.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/660.6/PRA/i/040902896 |
| Subjects: | 600 Technology (Applied sciences) > 660 Chemical engineering and related technologies > 660.6 Biotechnology |
| Divisions: | S2/S3 > Magister Matematika, Fakultas MIPA |
| Depositing User: | Endro Setyobudi |
| Date Deposited: | 19 Mar 2014 11:21 |
| Last Modified: | 19 Mar 2014 11:21 |
| URI: | http://repository.ub.ac.id/id/eprint/160095 |
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