Sains Malaysiana 38(2): 261-269(2009)

Pengesanan Gen Col1a2 Odontoblas Pada Tisu Pulpa

Gigi Oryctolagus Cuniculus

(Detection of CollA2 gene from Oryctolagus cuniculus Pulp Tissues)

Shahrul Hisham Zainal Ariffin, Noreena Nordin, Foo Siah Li,

Eni Juliana Ariffin, Sahidan Senafi

Pusat Pengajian Biosains dan Bioteknologi, Fakulti Sains dan Teknologi

Universiti Kebangsaan Malaysaia, Bangi, Selangor

Rohaya Megat Abdul Wahab

Jabatan Ortodontik, Fakulti Pergigian

Universiti Kebangsaan Malaysia, Kuala Lumpur

Received :  17 April 2008 / Accepted : 11 June 2008

ABSTRAK

 Kolagen jenis 1 α- 2 adalah salah satu daripada kolagen yang dapat ditemui dalam tisu pulpa gigi. Kolagen jenis 1 membentuk hampir 56% daripada kawasan intraselular di dalam pulpa. Objektif  kajian ini adalah untuk memencilkan gen Col1A2 daripada tisu pulpa Orycytolagus cuniculus. Gen penyelenggara, iaitu gen gliseraldehid 3-fosfat dehidrogenase (GAPDH) telah dipilih sebagai gen kawalan positif. Pencetus spesifik bagi kedua-dua gen Col1A2 dan GAPDH telah direkabentuk berdasarkan perisian Primer Premier Versi 5 berdasarkan jujukan yang diperolehi daripada NCBI (National Center for Biotechnology Information) RNA dipencilkan daripada tisu pulpa Oryctolagus cuniculus. Kedua-dua gen telah  diamplifikasi dan hasil produk PCR,  bersaiz 893 pb (Col1A2) dan  229 pb (GAPDH) telah didapati. Hasil analisis jujukan menunjukkan bahawa klon, Col1A2 mempunyai hampir 99% persamaan manakala GAPDH mempunyai 100% persamaan dengan jujukan yang terdapat dalam pengkalan data NCBI. Ini menunjukkan transkrip Col1A2 berjaya dikesan daripada tisu pulpa O. cuniculus dewasa.

Kata kunci : Odontoblas; dentin; tisu pulpa; Oryctolagus cuniculus; RT-PCR

ABSTRACT 

Type I collagen α-2 gene is a member of collagen group that can be found in pulp tissues. Type I collagen constitutes about 56% of large intercellular spaces in the pulp. The objective of this study is to isolate the Col1A2 gene, directly from RNA isolation from pulp tissue Orycytolagus cuniculus. A housekeeping gene, i.e., GAPDH is used as a positive control gene. Specific primer sets for Col12 and GAPDH genes have been designed using Primer Premier Version 5 based on both gene sequences obtained from NCBI (National Center for Biotechnology Information) [www.ncbi.nlm.nih.gov]. RNA had been isolated directly from pulp tissues of Oryctolagus cuniculus. The PCR products for Col1A2 and GAPDH were successfully amplified with the size of 893 bp (Col1A2) and 229 bp (GAPDH). Sequencing results had shown that Col1A2 and GAPDH clone attained almost 99% similarity and 100% identical compared to the sequence in the NCBI database respectively. This study shows that Col1A2 gene has been successfully detected from pulp tissue of O. cuniculus.

Keywords : Odontoblast; dentine; pulp Tissues; Oryctolagus cuniculus; RT-PCR

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