AMPLIFIKASI FRAGMEN PELACAK GEN LIPASE BAKTERI TERMOFILIK YANG DIISOLASI DARI KOMPOS

Agus Hery Susanto, Adi Amurwanto, Daniel Joko Wahyono, Saefuddin ’Aziz

Abstract


A study on amplification of a lipase gene probe of a thermophilic bacterial isolated from compost by the use of PCR technique was conducted at the Laboratory of Genetics, Biology Faculty, Jenderal Soedirman University, from October 2005 to January 2006. Bacillus sp strain ITB-A1, a thermophilic bacterial of compost origin showing lipase activity, was used in this study which could basically be divided into two main steps, i.e. chromosomal DNA isolation and in vitro amplification of a lipase gene probe. The primers used were upper primer (5’ GAG AGS RTG ATG AAA KGC TG 3’) and lower primer (5’ GAC AAG CAT GCG GGC CGT CTG 3’), respectively designed on the basis of conserved areas of lipase sequences of nine Bacillus genera and two Pseudomonas genera. Chromosomal DNA isolation indicated that the DNA was sufficiently good to be used as a PCR template due to no physical fragmentation observed. The PCR resulted in a thick band of approximately 500 bp, two thin bands assumed as a mispriming product, and a thin band assumed as a primer dimer. It could be concluded that an about 500 bp lipase gene probe of a thermophilic bacterial of compost origin was successfully in vitro amplified.

 

Keywords: thermophilic bacterial, compost, DNA probe, PCR


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