() | ||||
pAYC6 | RK2/RP4 (IncP) | Km | Methylomicrobium album BG8 (Berson and Lidstrom, 1997) | ???????? |
pK18mob | pBBR1 | Km | Methylosinus trichosporium OB3b (Stafford et al., 2003), Methylococcus capsulatus Bath (Csaki et al., 2003) | ?????? |
pCM184 | pBBR1 | Tc, Amp, Km | Methylocella sylvestris (Crombie and Murrell, 2014) Methylomicrobium alcaliphilum 20Z (Ojala et al.,2011 | ??????????? |
pSUKSM | RK2/RP4 (IncP) | Km | Methylomonas sp. st. 16a (Ye et al., 2007) | ?????????7 |
pJQ200SK | RSF1010 (IncQ) | Gm | Methylococcus capsulatus Bath (Welander and Summons, 2012) | ??????????????7 |
pCM433kanT | RK2/RP4 (IncP) | Km | Methylomicrobium buryatense (Puri et al., 2015) | ????????? |
1.3.
, , . , E.coli , , (Balasubramanian et al., 2010). , pMMO, . - pMMO E. coli, . , sMMO -. (Jahng et al., 1996; West et al., 1992). , - , . - E.coli 13- (Miller et al., 2015). -, . -, . , , , , .
, (, 2-3 Methylomonas .; Koffaset al., 2001)
|
|
, - , , (Kalyuzhnaya et al., 2013; Koffas et al.,2001). 64% (Haynes Gonzalez, 2014). , 2, . , , pMMO, .
. , , , - - (MCR) , -.
. Haynes and Gonzalez (2014).
, , .
1.4. , .
(Anthony, 1982; Trotsenko and Khmelenina, 2008;Smith and Murrell, 2009, 2010; Winder, 2010; Jiang et al., 2010; Feiet al., 2014). , . - ( ), , , . , (Egorov et al., 1990), (Nofferm/Calysta) Uniprotein (Unibio /). , / (http://www.unibio.dk, http://www.bioprotein. , http://www.calysta.com).
, ,: () , (Zimin, 2008); () , 2 (2 H4); () ( 10% ) , (Yazdian et al., 2005); (D) (Khmelenina et al., 1992); () , - (-: ) ; () . (Adedayo et al., 2011). , (Bothe et al., 2002) , , , , , , , .
|
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, : (3- (), (), (Xin et al., 2004a,b; Park and Lee, 2013); , , , (Trotsenko and Khmelenina,2008), / (Choi et al., 2011; Fei et al., 2014), (Koffas et al., 2005; Ye and Kelly,2012).
II , (Hou,1984) , -β- () (Zhang et al., 2009).
, -β- 50% (Doronina et al., 2008; Criddle et al., 2013).
-β- , .
(Corder et al., 1988; Labinger, 1995; Xin et al., 2004a,b; Cantrell et al., 2008; Park and Lee, 2013). II ,
M. Trichosporium, , , 2 MeDH (Mehta et al., 1991; Furuto et al.,1999; Xin et al., 2004a)
(Xin et al.,2004b)
, , .
.
: (Streese and Stegmann,2003; Yoon et al., 2009; Huang et al., 2011), (Semrau, 2011) (DiSpirito et al., 2011)
- , , , -, , -, - - , , (Semrau, 2011).
/ (Guiot et al., 2008).
(Park et al., 2008;Scheutz et al., 2009)
, (pMM), chalkophore, (), (10-21) (El Ghazouani et al., 2011; Kim et al., 2004)
, (Summeret al., 2011; Zischka et al., 2011).
, , , , (Choi et al., 2006).
|
|
.
, (Choi et al.,2006; DiSpirito et al., 2011; Vorobev et al., 2013).
, (Semrau et al., 2013)
, , , , , .
2. .
. .
2.1.
4 , Whittenbury et al. (1970)
22 (. www. methanotroph.org 2.1 ).
, , , ; .
, . . - ( ), , . . - , , . (Semrau et al., 2011; Dunfield and Dedysh, 2014)
- , .
Dedysh Dunfield (2011) , , , - , - .
.
, , , ( ), .
, () , , Methylococcus capsulatus Bath, M. trichosporium OB3b and Methylocystis spp.
, , .
, , , ; , (Kalyuzhnaya et al., 1999; Trotsenko and Murrell, 2008; Murrell,2010).
(1-10), ( 4 65 1) (0-10%) (Trotsenko and Khmelenina, 2008)
|
|
, ( NC10) (Ettwig et al., 2009)
pMMO, NC10 , N- 2.
2.2.
, (. www.methanotroph.org 5.1).
( ), .
- , , , , .
( , , , ) (Kao et al., 2004; Murrell and Jetten, 2009; Khadem et al., 2011;Campbell et al., 2011; Yang et al., 2013; Kalyuzhnaya et al., 2013;Vorobev et al., 2014; Tamas et al., 2014).
3.
, . - . , .
3.1.
RK2 / RP4 .
, , , , pRK2013 (Figurski Helinski, 1979)
Triparental -, , , pRK2013
( , 1979).
s17-1 , , , ( .,. 1983).
, methanotroph, () , , , , .
, .
25 / , - .
- .
.
PMO Methylocystis sp. st. SC2 (Baani and Liesack, 2008).
. silvstris bl2 . ( , 2011).
, .
3.2. / ()
, , , 500 .
|
|
, pCM184, , loxP, Cre (Marxand Lidstrom, 2002).
, .
sacB , M. capsulatus Bath,Methylomonas sp. st. 16a, and Methylomicrobium buryatense (Yeet al., 2007; Welander and Summons, 2012; Puri et al., 2015).
( ) .
tn5, Methylosinus sp. st. 6, . (Toukdarian Lidstrom, 1984).
.
, tn5 , , Methylomonassp. st. 16a (Sharpe et al., 2007).
3.3.
.
IncP , , pBBR IncQ , ( 1).
, .
GFP, dTomato, ( xylE) - ( LacZ) , , (Ali and Murrell, 2009; Ojala et al.,2011; Puri et al., 2015).
, .
4.
, ; , , , (Lee et al., 2008; Zelle et al.,2008; Lee et al., 2012; Karr et al. 2012; Esvelt and Wang, 2013).
. 1-, (VanDienandLidstrom, 2002; Peyraud et al., 2011).
, (Sipkema et al., 2000; Yoon and Semrau, 2008)
, 1, , , (Leak and Dalton, 1986)
" " (Santos et al., 2011).
, BioCyc (http://www.biocyc.org).
, , ( , , , /) , , COBRA (http://opencobra.sourceforge.net/open COBRA/Welcome.html) PathwayTools (bioinformatics.ai.sri.com/ ptools). , .
: (, - ), , ICMs,, .
GSM-FBA , , capsulatus . . trichosporium, , .
5.
. I - II , , (. 3).
, I , , , (. 3) ( ), , (Eshinimaev ., 2002; Koffas ., 2005). .
5.1. -.
, -- , .
(. 4).
EPS / I , -, .
, , (Khmelenina . 1992).
- II , PHB. , 2, , - , , 2 ( ., 2013).
, , . H2- , . , , , , 2 . , N2 , 2- .
I . , I ( . 2013). , 1 nadh 1.7 3 . , , , . - - , 1- ()
- I , CO2. , - ( ., 2013) , - RuMP (. 4). - RuMP . , RuMP , , .
, - , , , , , .
5.2. 2-
I , , 2 .
2 , 2- , .
, , .
, , .
, , .
5.3.
, - , pMMO (, 1982; Semrau ., 2010).
, , , .
I , II .
, , , , E. coli.
tesA ( ., 2010).
Synechocystis, , ( .,. 2011).
, atfA , ( Steinbuchel, 2014; ,. 2010). , , () ( .,1992). , atfA FAME ( 2010,).
- .