Transposon Insertion Phenomenon during Cloning of a Partial Fragment Derived from Metagenomic DNA Isolated from Deep-Sea Water and Sediment of Kawio Island, North Sulawesi
Abstract: Transposon is
well-known as mobile element found abundant both in prokaryote and eukaryote
genomes. In bacteria, transposon (famous name of a transposable DNA) could jump
from chromosome to plasmid and its contrary. One type of transposons in
bacteria known as insertion sequence (IS), it does not contain any additional
genes except a gene encoding transposase, an enzyme that correlated to
transponsition activities. The finding of transposon insertion unfortunately
found during cloning of a fragment derived from deep-sea metagenomic DNA in
this research. In the initial, this research was aimed to clone and
characterize the รก-amylase encoded gene derived from metagenomic DNA isolated
from deep-sea water and sediment of Kawio Island, North Sulawesi. Metagenomic
DNA has been isolated from deep-sea water and sediment and by using Whole
Genome Amplification (WGA) technique, the DNA it could be increased in
quantities to 146,31 ng for each 1 ng of metagenomic DNA. A fragment of ~1000
bp in length was obtained by using touchdown PCR method. The presence of a
transposon in this DNA fragment is proposed as a hypothesis for losing ~700 bp
leaving just 310 bp cloned sequence. Analysis of sequencing result showed a
highest similarity between this 310 bp partial fragment with a replication
protein (Rep) encoded gene from Pseudomonas putida (Query Coverage: 88%; Max.
Identity: 80%, Positive: 86%) and this protein is known to be involved in
plasmid replication where transposase encoding genes known usually presence
together with this gene (Rep gene) in a bacterial plasmid.
Keywords: transposon,
metagenomic; partial fragment; cloning; deep-sea
Author: Mochamad Untung Kurnia
Agung, Maelita Ramdani Moeis
Journal Code: jpperikanangg130017
