TABLE 1.
Phenotypic screening of strains with plasmid-carried tcpB
| Construct | Alteration(s)a | TcpB stabilityb | Autoagglutinationb | Transduction efficiencyc | TcpF secretionb |
|---|---|---|---|---|---|
| pTK85 | WT TcpB | + | + | ++ | + |
| pBAD22 | Empty vector | − | − | − | − |
| pJMA82 | E5V | + | + | ++ | − |
| pJMA83 | R26E | + | + | + | + |
| pJMA86 | W79A | + | + | ++ | +/− |
| pJMA85 | K81A | + | + | ++ | + |
| pJMA84 | E83R | + | + | ++ | + |
| pJMA37 | C85A | − | − | ++ | +/− |
| pJMA72 | C85S | − | − | − | − |
| pJMA54 | C107A | − | − | + | + |
| pJMA73 | C107S | + | − | + | − |
| pJMA45 | LCWD249-252AAAA | +/− | − | +/− | − |
| pCHG016 | L249A | + | + | ++ | +/− |
| pJMA38 | C250A | + | − | − | +/− |
| pJMA75 | C250S | − | − | − | − |
| pCHG017 | W251A | + | − | ++ | − |
| pJMA58 | D252A | +/− | + | ++ | + |
| pJMA39 | C261A | + | − | +/− | +/− |
| pJMA76 | C261S | + | − | +/− | − |
| pJMA52 | K272A | + | + | ++ | + |
| pJMA62 | E274A | + | + | ++ | + |
| pJMA61 | D276A | + | + | ++ | + |
| pJMA49 | K278A | + | + | ++ | +/− |
| pJMA51 | D281A | +/− | + | ++ | +/− |
| pJMA60 | K286A | + | + | ++ | + |
| pJMA48 | K295A | + | + | ++ | + |
| pJMA53 | K297A | + | + | ++ | + |
| pJMA69 | F307A | + | − | + | +/− |
| pJMA47 | D309A | + | + | ++ | + |
| pJMA50 | K314A | +/− | ++ | ++ | + |
| pJMA40 | C321A | − | − | − | +/− |
| pJMA74 | C321S | +/− | − | ++ | − |
| pJMA41 | R328A | + | + | ++ | + |
| pJMA68 | SS334-335AA | + | − | + | +/− |
| pJMA42 | E348A | + | + | ++ | + |
| pJMA34 | KD351-2AA | +/− | − | ++ | − |
| pJMA46 | L262P, KD351-2AA | − | +/− | ++ | − |
| pJMA67 | S353A | + | +/− | + | +/− |
| pJMA35 | KD359-60AA | + | + | ++ | + |
| pJMA66 | S362A | + | +/− | ++ | + |
| pJMA32 | K363A | + | + | ++ | + |
| pJMA59 | H366A | + | + | ++ | +/− |
| pJMA65 | L369A | + | + | ++ | + |
| pJMA64 | S370A | + | + | ++ | + |
| pJMA43 | K381A | + | + | ++ | + |
| pJMA63 | W383A | + | − | + | +/− |
| pJMA33 | D384A | + | + | ++ | + |
| pJMA55 | E392A | + | +/− | ++ | + |
| pJMA70 | E392A, D402A | +/− | + | + | +/− |
| pJMA56 | D402A | + | + | ++ | + |
| pJMA36 | RNPK408-11AAAA | + | − | − | +/− |
| pJMA57 | RNPK408-11ANPA | + | + | ++ | + |
| pJMA71 | Δ392-401, RNPK408-11AKNA | +/− | − | − | + |
| pCHG005 | W420A | − | − | − | − |
| pCHG003 | C421S | − | − | − | − |
a
Alterations in bold type were selected for further study as chromosomal mutations.
b
+, wild type like; −, ΔtcpB mutant like; +/−, a decrease in function compared to the wild type but not complete loss; ++, exceeds wild-type levels.
c
Level of transduction efficiency: ++, approximately wild-type level; +, approximately 1 to 2 log below wild-type level; +/− approximately 3 to 5 log below wild-type level; −, no Kmr bacteria were recovered and transduction efficiency is therefore below the limit of detection, equivalent to >7 to 8 log below the wild-type level.