Natural competence in Thermoanaerobacter and Thermoanaerobacterium species.
Low-G+C thermophilic obligate anaerobes in the class Clostridia are considered among the bacteria most resistant to genetic engineering due to the difficulty of introducing foreign DNA, thus limiting the ability to study and exploit their native hydrolytic and fermentative capabilities. Here, we report evidence of natural genetic competence in 13 Thermoanaerobacter and Thermoanaerobacterium strains previously believed to be difficult to transform or genetically recalcitrant. In Thermoanaerobacterium saccharolyticum JW/SL-YS485, natural competence-mediated DNA incorporation occurs during the exponential growth phase with both replicating plasmid and homologous recombination-based integration, and circular or linear DNA. In T. saccharolyticum, disruptions of genes similar to comEA, comEC, and a type IV pilus (T4P) gene operon result in strains unable to incorporate further DNA, suggesting that natural competence occurs via a conserved Gram-positive mechanism. The relative ease of employing natural competence for gene transfer should foster genetic engineering in these industrially relevant organisms, and understanding the mechanisms underlying natural competence may be useful in increasing the applicability of genetic tools to difficult-to-transform organisms.
Shaw
A Joe
AJ
Mascoma Corporation, Lebanon, NH 037661, USA.
Hogsett
David A
DA
Lynd
Lee R
LR
D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004270:DNA, Circular / Q000378:metabolism; D017353:Gene Deletion; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010957:Plasmids; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D046968:Thermoanaerobacter / Q000235:genetics*; D046950:Thermoanaerobacterium / Q000235:genetics*; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular
Biotechnol Prog. 2001 Jan-Feb;17(1):52-6
11170479
Mol Microbiol. 2002 Oct;46(1):25-36
12366828
Arch Microbiol. 1984 May;138(1):31-6
6742954
Science. 2008 Dec 19;322(5909):1843-5
19095942
Arch Microbiol. 1997 Aug;168(2):114-9
9238102
Annu Rev Microbiol. 1986;40:211-35
3535646
J Bacteriol. 1986 Apr;166(1):338-40
3957870
Appl Environ Microbiol. 2008 Nov;74(21):6720-9
18776029
Appl Environ Microbiol. 1985 Mar;49(3):656-9
16346758
Appl Environ Microbiol. 2006 Jul;72(7):5027-36
16820502
Nucleic Acids Res. 2004 Oct 28;32(19):5780-90
15514111
Curr Opin Biotechnol. 2005 Oct;16(5):577-83
16154338
Nat Rev Microbiol. 2004 Mar;2(3):241-9
15083159
Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11140-4
7479953
J Biol Chem. 2006 Aug 4;281(31):21720-21727
16751195
Syst Appl Microbiol. 2005 Aug;28(6):555-61
16104353
Int J Syst Bacteriol. 1995 Oct;45(4):783-9
7547300
Biochem Soc Trans. 2004 Apr;32(Pt 2):283-9
15046590
Appl Environ Microbiol. 1995 Mar;61(3):937-40
16534977
Trends Biotechnol. 2009 Jul;27(7):398-405
19481826
J Bacteriol. 1979 Sep;139(3):800-10
39062
Appl Environ Microbiol. 1991 Feb;57(2):579-85
16348422
Res Microbiol. 2007 Dec;158(10):767-78
17997281
Mol Microbiol. 1992 May;6(10):1263-70
1640829
Proc Natl Acad Sci U S A. 2008 Sep 16;105(37):13769-74
18779592
J Bacteriol. 2009 Oct;191(20):6457-64
19648238
Microbiol Mol Biol Rev. 2002 Sep;66(3):506-77, table of contents
12209002
Microbiol Rev. 1994 Sep;58(3):563-602
7968924
Mol Microbiol. 1998 Feb;27(4):845-52
9515709
Microbiology (Reading). 2009 Mar;155(Pt 3):852-862
19246756
Int J Syst Evol Microbiol. 2004 Mar;54(Pt 2):467-474
15023962
Biotechnol Prog. 1998 Jan-Feb;14(1):116-25
9496676
Appl Environ Microbiol. 2005 Dec;71(12):8069-76
16332787
Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7986-90
7644525
Anaerobe. 2006 Jun;12(3):153-9
16698292
Mol Microbiol. 2003 Oct;50(2):527-36
14617176
Appl Environ Microbiol. 2002 Dec;68(12):6013-20
12450823
Microbiol Mol Biol Rev. 2005 Mar;69(1):124-54
15755956
Nat Rev Microbiol. 2009 Oct;7(10):715-23
19756010
Appl Microbiol Biotechnol. 2007 Nov;77(1):61-8
17899073
Appl Microbiol Biotechnol. 2004 Oct;65(5):600-5
15007569
J Gen Microbiol. 1993 Feb;139(2):295-305
8436948
Microbiol Rev. 1993 Jun;57(2):451-509
8336675