Conjugation

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Schematic diagram of bacterial conjugation. 1 - Donor cell produces pilus. 2 - Pilus attaches to recipient cell, brings the two cells together. 3 - The mobile plasmid is nicked and a single strand of DNA is then transferred to the recipient cell. 4 - Both cells recircularize their plasmids, synthesize second strands, and reproduce pili; both cells are now viable donors. Image courtesy of [http://en.wikipedia.org/wiki/Bacterial_conjugation Wikipedia]

Bacterial conjugation is the transfer of genetic material between two bacterial cell via direct cell-to-cell contact. Although often incorrectly characterized as the bacterial equivalent of mating, in fact it is simply the transfer of genetic material from a donor cell to a recipient cell. Bacterial conjugation is also known as a "type IV secretion system".

This behaviour is characterized by the presence of a specialized plasmid (small, circular piece of transferrable DNA most often found within bacteria) known as a conjugative plasmid. The conjugative plasmid holds a distinct and specialized set of coding regions including the following

• OriT (Origin of Transfer): Unlike other plasmids, the conjugative plasmid has its own origin of transfer nic region. This is a specialized portion of the genome where one of the two strands of the circular plasmid DNA is cut to allow for rolling circle replication of the plasmid containing the OriT region into a recipient cell. During rolling circle replication, the cut end of one strand is inserted into the recipient cell while recipient polymerases construct the complementary strand back into a circular piece of DNA.
• TraJ (Transfer genes): The activation of this gene sets off a cascade of other plasmid genes (and thus corresponding protein expressions) which act in concert to form "mating" characteristics in the host bacterial cell. Such salient characteristics include:
  • growth of a pilus--a long whip-like apparatus used to hook the two cells together
  • fusion of the outer membranes to allow for transfer of genetic material
  • formation of "surface exclusion proteins" which prevents the cell containing the conjugative plasmid from mating with other conjugative plasmids of its type

Two of the most well-studied and characterized types of conjugative plasmids are types "F" (from the F plasmid) and "R". The employed nomenclature for conjugative plasmid genes is the gene name first with the type afterwards. For example, the "TraJr" or "TraJR" gene denotes the "TraJ" gene of the "R" type conjugative plasmid.

Note it was previously believed that transfer of genetic material occurred via the pilus itself; now, most think that the transfer of genetic material occurs via a separate direct channel between the two cells.

More...

Name	Type	Description	Length
BBa_I714000	Conjugation	R388 oriT ( transfer origin of E.coli IncW group R388 plasmid)	402
BBa_I714003	Conjugation	minimum oriT of plasmid R64 ( origin of transfer, R64 is IncI-1 )	83
BBa_I714030	Conjugation	OriT-F (Origin of Transfer for the F-plasmid nic region)	355
BBa_I714031	Conjugation	OriT-R (Origin of Transfer for the R751-plasmid nic region)	278
BBa_I714032	Conjugation	OriT-S (Origin of Transfer for the pSC101-plasmid nic region)	131
BBa_J01000	Coding	TraJF controls conjugative transfer in F plasmid	690
BBa_J01001	Coding	TraJr protein (controls conjugative transfer in RP4 plasmid)	378
BBa_J01002	Conjugation	OriT-F (Origin of Transfer for the F-plasmid nic region)	365
BBa_J01003	Conjugation	OriT-R (Origin of transfer for the R-plasmid nic region)	371
BBa_J01077	Plasmid	F plasmid (x-OriT, x-TraJ)
BBa_J01078	Plasmid	R plasmid (x-OriT, x-TraJ)
BBa_J107185	Conjugation	oriT from RK2	354
BBa_J23046	Conjugation	put in later - SIL strain
BBa_J61030	Composite	[OriTr][R6K]	785
BBa_K1053123	Conjugation	Split EYFP N-terminal domain fused with HTLV-1 Rex peptide	606
BBa_K1053124	Conjugation	Split EYFP C-terminal domain fused with λN peptide	297
BBa_K1084999	Conjugation	GGA_vector	125
BBa_K109550	Coding	Bacterial conjugative regulator KorA	306
BBa_K109551	Coding	TrbA transcription factor for the Tra2 operon in incP alpha conjugation	366
BBa_K125320	Conjugation	OriT-RP4 (Origin of transfer for the RP4 plasmid)	99
BBa_K125350	Conjugation	oriTr with lac promoter and lacZ	979
BBa_K125360	Conjugation	oriT with lac promoter and lacZ	707
BBa_K1439000	Conjugation	Origin of transfer for the RP4-plasmid nic region.	350
BBa_K150006	Conjugation	oriT	471
BBa_K1510119	Conjugation	A truncated INPNC anchor protein that binds fragments on the cell surface	1564
BBa_K1510214	Conjugation	Dispersin generator circuit	2053
BBa_K1510409	Conjugation	A Steptococcus Mutants adhesive complex.	1780
BBa_K1510603	Conjugation	Dispersin generator ,with constitutive promoter plus yebf	1916
BBa_K1510624	Conjugation	Truncated lysostaphin coding sequence plus strong promoter and yebf	1331
BBa_K1510816	Conjugation	A Steptococcus Mutants adhesive complex.	1651
BBa_K1510831	Conjugation	Fragrance generator part2: alcohol acetyltransferase I and RBS	1599
BBa_K1510904	Conjugation	Truncated lysostaphin coding circuit	1468
BBa_K1510915	Conjugation	ketoisocaproate decarboxylase (THI3) Circuit	1985
BBa_K1510922	Conjugation	Fragrance generator part1: ketoisocaproate decarboxylase (THI3) and RBS	1848
BBa_K1510925	Conjugation	Fragrance generator3: branched-chain amino acid transaminase (BAT2) and RBS	1152
BBa_K1682014	Conjugation	Conjugation(RP4) plasmid with partial GFP	1395
BBa_K175000	Coding	trbC (pilin) of IncP beta R751 plasmid	468
BBa_K175001	Coding	trbK (entry exclusion) of IncP beta R751 plasmid	231
BBa_K188777	Conjugation	aka	879
BBa_K188778	Conjugation	as	864
BBa_K2144101	Plasmid_Backbone	Vector containing Hisx6 and Sortase tag LPETGG	2117
BBa_K2657002	Conjugation	Rp4 Origin of Transfer PhytoBrick	131
BBa_K2745002	Conjugation	linker	15
BBa_K2759003	Conjugation	Bacterioferritin (BFR) with sfGFP conjugation	1203
BBa_K2791011	Conjugation	Translational coupling device, weak.	32
BBa_K2791012	Conjugation	Translational coupling device, strong.	32
BBa_K2791013	Conjugation	Translational coupling device, medium.	32
BBa_K2791014	Conjugation	Translational coupling device, very weak.	32
BBa_K2796006	Conjugation	P2A	66
BBa_K2796007	Conjugation	T2A	63
BBa_K2913002	Conjugation	A conjugation between Pluc anad HucO	11
BBa_K2913008	Conjugation	A referenced conjugation between Pluc and CDS sequence	35
BBa_K2961003	Conjugation	gRNA / crRNA fixed-loop repeat for Cpf1 / Cas12a	21
BBa_K2976010	Conjugation	Improved T2A	63
BBa_K2976011	Conjugation	(Gly4Ser)3 linker	30
BBa_K2976012	Conjugation	Gly-Ser-Ser linker	9
BBa_K3190206	Conjugation	Linker	24
BBa_K3819025	Conjugation	Pro/Thr-rich Linker	108
BBa_K3875021	Conjugation	Cas9 binding	34
BBa_K3946006	Conjugation	L2NC silica tag+term (CT)	4461
BBa_K3946007	Conjugation	L2NC+linker+term (CT)	4516
BBa_K3946044	Conjugation	Sb7 silica tag (C)-PETase (W159H/ S238F)	902
BBa_K3946045	Conjugation	Sb7 silica tag (C)-LCC-WCCG variant	811
BBa_K4103001	Conjugation	pIP501 transfer origin	378
BBa_K4103016	Conjugation	5'lacA	508
BBa_K4122007	Conjugation	spytag
BBa_K4122008	Conjugation	Spycatcher
BBa_K4122009	Conjugation	Snooptag
BBa_K4122010	Conjugation	Snoopcatcher
BBa_K4170009	Conjugation	Serine-serine-glycine flexible linker	9
BBa_K4580004	Conjugation	NdmB-6x his-GFP	1806
BBa_K4587106	Conjugation	GS linker	48
BBa_K4643003	Conjugation	incP origin of transfer	109
BBa_K4651000	Conjugation	sdjhf
BBa_K4696019	Conjugation	P2A	57
BBa_K4761030	Conjugation	linker1	15
BBa_K4761031	Conjugation	linker2	45
BBa_K4761032	Conjugation	linker3	24
BBa_K4761033	Conjugation	linker4	15
BBa_K4761034	Conjugation	linker5	45
BBa_K4761035	Conjugation	linker6	15
BBa_K4765023	RNA	PmeI linker	27
BBa_K4971011	Conjugation	Linker protein sequence with starting codon	30
BBa_K5096400	Conjugation	pCas9	8437
BBa_K5170000	Conjugation	A novel PROTAC using BBa_K5170004 against 231pTau	66
BBa_K5170001	Conjugation	A novel PROTAC using BBa_K5170002 against 231pTau	66
BBa_K5170005	Conjugation	A novel PROTAC using BBa_K5170003 against 231pTau	66
BBa_K543016	Conjugation	Bacteriophage Lysis Cassette S105, R, and Rz under Tet promoter (conjugative)	1838
BBa_K543020	Conjugation	OriTr-GFP	1254
BBa_K543030	Conjugation	[I714030][J23066][I714030] (Constitutive expression)	1105
BBa_K543032	Conjugation	assay system of killing by conjugation	3615
BBa_K5436020	Conjugation	BamHI Linker	6
BBa_K5465999	Plasmid	ONERING-TetO	6902
BBa_K640002	Conjugation	oriT - Interspecies origin of transfer	260
BBa_K640004	Conjugation	Device for conjugation of plasmids from E. coli to pseudomonas	642
BBa_K640005	Conjugation	Device for conjugating plasmids from E. coli to pseudomonas	753
BBa_K808028	Signalling	PhoA: signal sequence for peri plasmic expression	63
BBa_K873003	Conjugation	conjugative transfer of RFG	1302
BBa_M1355	Conjugation	RS1	720
BBa_M1356	Conjugation	RS2	709

Yifan Yang, Mingzhi Qu and the 2007 Peking Yniveristy iGEM team designed the parts BBa_I714000, BBa_I714003, BBa_I714030, BBa_I714031 and BBa_I714032.

The 2005 Berkeley iGEM team designed the parts BBa_J01000-BBa_J01003 and BBa_J01077-BBa_J01078.

Chris Anderson and the 2006 Berkeley iGEM team design the parts BBa_J23046.

Ingrid Swanson and the 2008 University of Washington iGEM team designed the parts BBa_K109550 and BBa_K109551.

Margaret Ruzicki and the 2008 University of Hawaii iGEM team designed the parts Part:BBa_K125320, Part:BBa_K125350, and Part:BBa_K125360.

References

<biblio>

1. Frost pmid=7915817
2. Jaenecke pmid=8858584
3. Lawley pmid=12855161
4. Lessl pmid=8407818
5. Miller pmid=2993231
6. Martinez-Morales pmid=10559184
7. Wilkins pmid=12220405

</biblio>

• [http://en.wikipedia.org/wiki/Bacterial_conjugation Wikipedia entry on Bacterial Conjugation]
• 2005 UC Berkeley iGEM team [http://2006.igem.org/Berkeley_2005 project] and [http://2006.igem.org/Berkeley_Protocols protocols]