E. coli J53 is always used as a recipient strain, but could it conjugate later?
if the plasmid is selftransferable -yes, if the strain contains tra genes can mobilize plasmids too, i do not know this particular strain.
J53 is an F+ strain, meaning that it carries the F plasmid which can conjugate. So yes it can conjugate later, however whether it will conjugate the particular plasmid you are using depends upon the plasmid itself and whether or not F conjugation system will work on it or not.
How can we distinguish between E.coli lab. strains properties to set them as donor/recipients?
For ex;
Can we use E.coli J53 (3876) with RP4 plasmid as donor and
the rifampycin resistant E.coli J53 as recipient?
What i have came along in literature so far is HT99 as donor and Rifampycin res. E.coli J53 as recipient.
I did a bit more research in the literature and need to amend my earlier answer to this question. The original J5-3 strain is F+ and therefore is capable of conjugation. It was subsequently cured of F and the new F- strain is called J53 (instead of J5-3). In various places in the literature they get mixed so for example Ecoliwiki list J53 as F+. Therefore the best answer for those of you out there is you need to either test your strain for whether it carries F (the simplest test is whether M13 forms plaques or not, although certainly you can do PCR test for any of the F genes). Similarly, the most predominantly cited J53 strain is F- azi-R (azide resistant). It may well be that others have since made it Rif-R as well.
So my answer to Ilana Lopes Baratella da Cunha Camargo needs to be modified and it depends upon which J53 strain you have as to whether it can subsequently mobilize your plasmid.
My answer to Can Burak Ozkal is that you should be sure to check your strain is truly Rif-R before using it. However RP4 and F and entirely different in their transmissibility, so you can mobile an RP4 plasmid into J53 (any variety) and if you have J53(RP4) you can mobilize the RP4 to a different recipient as well.
As to your general question of how to distinguish whether an E. coli strain can serve as a donor or recipient, the simple (and true most of the time but not absolutely always) is that conjugative plasmids like RP4 can be mated to virtually any E. coli strain. And any strain that carries the plasmid can serve as a donor. The plasmid itself carries the genes for conjugation and makes the strain a suitable donor. The only requirement is that you have appropriate selectable markers so you can distinguish between the strains.
The above paragraph is true for wild type conjugative plasmids like F, RP4 and many others. However when you start talking about engineered plasmids in the laboratory then it becomes more complicated because many of the engineered plasmids have had the conjugation genes removed and require a helper plasmid in the strain to encode the conjugative functions.
Appreciate the very descriptive answer of yours Michael J. Benedik
Otherwise, It would be impractical to interpret (by simple literature research) the points you have highlighted and clarified.
I’m just making an experimental plan;
And planning to purchase the two engineered strains that i have recently mentioned.
So in practice is it possible that i may purchase a conjugative gene removed J53-Rp4 strain and a false J53-Rifampycin strain?
On which points i may pay attention and make a possible reminder to my supplier.
Thank you.
I think if you are purchasing the strains then they are probably going to be fine and what you expect. I'm sure the supplier has checked the essential features. The problem arises when you get a strain from a another lab and they may not know the history.
I need J53 azide resiatant strain. Can somebody provide me the same@ [email protected] . I require this for plasmid based Antimicrobial sensitivity transfer assay.
Regards
@MICHAELJ.BENEDIK thank you for your answer. If I have a F- rifR and use it to receive a conjugative plasmid (with all conjugation genes) then it will be able to conjugate, right? In my case, I want to select conjugative plasmids from clinical isolates, before sequencing... I understand that some plasmids need the presence of a helper plasmid, otherwise they won't conjugate. However, once those "complete" plasmid or a couple of plasmids that help each other are transfered to E. Coli J53, even if it is F-, they will be able to conjugate to a next isolate, right?
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