CLONING AND SEQUENCING OF GENE ENCODING β-GALACTOSIDASE OF BACTERIA ARTHROBACTER SULFONIVORANS
Abstract
A full nucleotide sequence of the β-galactosidase gene (β-gal) was deciphered by cloning in plasmid pJET1.2 of DNA fragments containing β-gal of bacteria Arthrobacter sulfonivorans LF-GAL, their subsequent sequencing and comparison with the known sequences from the GenBank database. It was deposited under access number KM2778940.
The evaluation of the β-gal structure showed that it is composed of 3132 b.p. encoding 1043 amino acids making up a β-galactosidase subunit with a molecular weight of 113.6 kDa. It was found that β-gal of the examined strain is characterized by the highest degree of similarity to the genes encoding β-galactosidase in the genus Arthrobacter (66–72 %).
The amino acid composition of enzyme proteins from A. sulfonivorans LF-GAL matches that of other representatives of the genus Arthrobacter by 59–95 %. The analysis of the enzyme protein amino acid sequence by BLAST software package demonstrated that β-galactosidase of A. sulfonivorans LF-GAL comprises conservative sequences typical for glycosyl hydrolase family 2.
About the Authors
A. A. KASTSIANEVICH
Институт микробиологии НАН Беларуси, Минск
Belarus
Belarus
L. I. SAPUNOVA
Институт микробиологии НАН Беларуси, Минск
Belarus
Belarus
A. G. LOBANOK
Институт микробиологии НАН Беларуси, Минск
Belarus
Belarus
References
1. Nath A., Mondal S., Chakraborty S. et al. // Asia-Pac. J. Chem. Eng. 2014. Vol. 9, is. 3. P. 330–348.
2. Panesar P. S., Kumari S., Panesar R. // Enzyme Res. 2010. Vol. 2010. P. 1–16.
3. Husain Q. // Crit. Rev. Biotechnol. 2010. Vol. 30, N 1. P. 41–62.
4. Tang L., Li Z. A., Dong X. X. // J. Ind. Microbiol. Biotechnol. 2011. Vol. 38. P. 471–476.
5. Asraf S. S., Gunasekaran P. // Current trends of ß-galactosidase research and application. Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology. Microbiology book series. 2010, N 2. P. 880–890.
6. Kastsianevich A. A., Sapunova L. I. // Сб. мат. II Междунар. науч.-практ. конф. молодых ученых «Научные стремления–2011». 2011. Т. 1. С. 200–206.
7. Сапунова Л. И., Костеневич А. А., Лобанок А. Г. и др. // Докл. НАН Беларуси. 2013. Т. 57, № 6. С. 70–74.
8. Сапунова Л. И., Костеневич А. А., Лобанок А. Г. и др. // Докл. НАН Беларуси. 2014. Т. 58, № 4. С. 71–77.
9. Способ получения внеклеточной β-галактозидазы. Патент 15050 РБ. а 20091062; заявл. 14.07.2009; опубл. 28.02.2011.
10. Сапунова Л. И., Костеневич А. А., Лобанок А. Г. и др. // Тр. БГУ, сер. физиол., биохимия и молекул. основы функционирования биосистем. 2013. Т. 8, Ч. 1. С. 224–229.
11. Костеневич А. А., Сапунова Л. И., Лобанок А. Г. // Микробные биотехнологии: фундаментальные и прикладные аспекты. 2014. Т. 6. С. 51–62.
12. Sambrook J., Frittsch E., Maniatis T. // Molecular cloning: a laboratory manual. 2nd ed. New York: Cold Spring Harbor Laboratory Press, 1989. – 2222 p.
13. http://molbiol.ru/protocol/03_05.html#a1
14. http://www.bioinformatics.babraham.ac.uk/projects/fastqc/
15. Bolger A. M., Lohse M., Usadel B. // Bioinformatics. 2014. Vol. 30. P. 2114–2120.
16. Bankevich A., Nurk S., Antipov D. // J. Comp. Biol. 2012. Vol. 19, N 5. P. 455–477.
17. Langmead B., Salzberg S. // Nature Methods. 2012. Vol. 9. P. 357–359.
18. Van Domselaar G. H., Stothard P., Shrivastava S. et al. // Nucleic Acids Res. 2005. Vol. 33. P. W455–459.
19. Altschul S. F., Gish W., Miller W. J. Mol. Biol. 1990. Vol. 215, N 3. P. 403–410.
20. http://www.ncbi.nlm.nih.gov/refseq/
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