TY - JOUR
T1 - Isolation of an erythromycin-resistant (putative) gene from bacillus halodurans CM1
AU - Widamayanti,
AU - Haniyya,
AU - Helianti, I.
AU - Abinawanto,
N1 - Funding Information:
This study was funded by the Insinas Research Incentive Program from theMinistry of Research, Technology and Higher Education 2019 granted to IH.
Publisher Copyright:
© the author(s)
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/10/7
Y1 - 2020/10/7
N2 - Bacillus halodurans CM1 is one of the bacterial strains from BPPT Culture Collection which had been isolated from sediments in the Cimanggu hot spring, West Java, previously. The whole genomic DNA of this strain has not yet been deeply studied despite its potential as host to produce several kinds of useful enzymes, such as xylanase, lipase, and protease. Besides the industrial enzymes, potential genes in B. halodurans CM1, for example antibiotic-resistant genes, could be engineered later for further purposes. Earlier experiments showed that CM1 strain has the ability to survive in the media with erythromycin, suggesting there might be genes responsible for this erythromycin resistance. This study aimed to isolate erythromycin-resistant gene from B. halodurans CM1 using a PCR approach. The primers used for PCR were designed based on the GenBank database of whole genome Bacillus halodurans C-125. The PCR product was then ligated into a pGEM-T easy cloning vector, and the recombinant vector then transformed into Escherichia coli DH5α. The DNA fragment with the size of 864 bp was obtained and it encoded 288 deduced amino acids. DNA sequence analysis showed that the gene had 99% similarity with gene encoding erythromycin resistance from B. halodurans C-125 (GenBank No access: BA000004.3, ErmK). There are 3 nucleotides in the ErmK sequence of B. halodurans CM1 that are different from that of another well-known strain B. halodurans C-125. This dissimilar pattern of sequence encodes Leu instead of Ser in 155th orders of CM1 amino acid sequence.
AB - Bacillus halodurans CM1 is one of the bacterial strains from BPPT Culture Collection which had been isolated from sediments in the Cimanggu hot spring, West Java, previously. The whole genomic DNA of this strain has not yet been deeply studied despite its potential as host to produce several kinds of useful enzymes, such as xylanase, lipase, and protease. Besides the industrial enzymes, potential genes in B. halodurans CM1, for example antibiotic-resistant genes, could be engineered later for further purposes. Earlier experiments showed that CM1 strain has the ability to survive in the media with erythromycin, suggesting there might be genes responsible for this erythromycin resistance. This study aimed to isolate erythromycin-resistant gene from B. halodurans CM1 using a PCR approach. The primers used for PCR were designed based on the GenBank database of whole genome Bacillus halodurans C-125. The PCR product was then ligated into a pGEM-T easy cloning vector, and the recombinant vector then transformed into Escherichia coli DH5α. The DNA fragment with the size of 864 bp was obtained and it encoded 288 deduced amino acids. DNA sequence analysis showed that the gene had 99% similarity with gene encoding erythromycin resistance from B. halodurans C-125 (GenBank No access: BA000004.3, ErmK). There are 3 nucleotides in the ErmK sequence of B. halodurans CM1 that are different from that of another well-known strain B. halodurans C-125. This dissimilar pattern of sequence encodes Leu instead of Ser in 155th orders of CM1 amino acid sequence.
UR - http://www.scopus.com/inward/record.url?scp=85092674946&partnerID=8YFLogxK
U2 - 10.1088/1755-1315/572/1/012017
DO - 10.1088/1755-1315/572/1/012017
M3 - Conference article
AN - SCOPUS:85092674946
SN - 1755-1307
VL - 572
JO - IOP Conference Series: Earth and Environmental Science
JF - IOP Conference Series: Earth and Environmental Science
IS - 1
M1 - 012017
T2 - 9th International Symposium for Sustainable Humanosphere, ISSH 2019
Y2 - 28 October 2019 through 29 October 2019
ER -