Boston University/Bacterial Conjugation Results
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===Filter conjugation to transfer the plasmid pMS291(lacZ) from E.coli SM10 to S. oneidensis AS-92=== | ===Filter conjugation to transfer the plasmid pMS291(lacZ) from E.coli SM10 to S. oneidensis AS-92=== | ||
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:The plasmid pMS291 is first transformed into E.coli SM10 and then the conjugation protocol is run to encourage conjugal transfer of the plasmid to S. oneidensis AS-92. A double-antibiotic selection method is used to select for successful S. onedensis transformants which have a resistance for both kanamycin and gentamicin. | :The plasmid pMS291 is first transformed into E.coli SM10 and then the conjugation protocol is run to encourage conjugal transfer of the plasmid to S. oneidensis AS-92. A double-antibiotic selection method is used to select for successful S. onedensis transformants which have a resistance for both kanamycin and gentamicin. | ||
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| + | [[Boston University/Conjugation | Read more about how conjugation works]] | ||
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| + | [[Boston University/Filter Conjugation Protocol | Our Protocol]] | ||
[[Image:BU_conjugation1.jpg]] | [[Image:BU_conjugation1.jpg]] | ||
Latest revision as of 16:41, 16 July 2007
Filter conjugation to transfer the plasmid pMS291(lacZ) from E.coli SM10 to S. oneidensis AS-92
- The plasmid pMS291 contained a kanamycin resistance gene. S. oneidensis has a native gentamicin resistance.
- The plasmid pMS291 is first transformed into E.coli SM10 and then the conjugation protocol is run to encourage conjugal transfer of the plasmid to S. oneidensis AS-92. A double-antibiotic selection method is used to select for successful S. onedensis transformants which have a resistance for both kanamycin and gentamicin.
Read more about how conjugation works
Plate 1: 7 separate conjugation mixes were plated. The white is the filter upon which the bacteria conjugate. S. oneidensis is red.
Plate 2
Plate 3
Plate 4
Plate 5
