Thanks for all the answers! A bit of feedback for you:
1. It looks like pGreen is a plasmid for bacterium mediated plant transformation, so not necessarily a great choice for cloning a gene into lactobacillus. It's been about 8.5 years since I was actively doing molecular biology, so I am no longer facile with all the cartridges and restriction sites, but it looks to me like pGreen is designed for gene transfer, not expression.
2. According to the web site, pGreen confers kanamycin resistance, not ampicillin, so if your plates are coming up empty (and in molecular biology, whenever anything goes wrong, the result is the same--empty plates), possibly it's because you are indiscriminately killing everything with ampicillin.
3. BUT, I did a very quick web search and it looks like lactobacillus is not necessarily susceptible to ampicillin. Ampicillin does work on gram positive bacteria, but not every gram positive bacteria is susceptible to every such antibiotic. Cloning is almost always done with e. coli, since the characteristics of the host cell are generally not of interest at the endpoint of the experiment.
4. If the protocol you found for transforming lactobacillus is complicated, then it is undoubtedly because simpler protocols don't work.
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1. It looks like pGreen is a plasmid for bacterium mediated plant transformation, so not necessarily a great choice for cloning a gene into lactobacillus. It's been about 8.5 years since I was actively doing molecular biology, so I am no longer facile with all the cartridges and restriction sites, but it looks to me like pGreen is designed for gene transfer, not expression.
2. According to the web site, pGreen confers kanamycin resistance, not ampicillin, so if your plates are coming up empty (and in molecular biology, whenever anything goes wrong, the result is the same--empty plates), possibly it's because you are indiscriminately killing everything with ampicillin.
3. BUT, I did a very quick web search and it looks like lactobacillus is not necessarily susceptible to ampicillin. Ampicillin does work on gram positive bacteria, but not every gram positive bacteria is susceptible to every such antibiotic. Cloning is almost always done with e. coli, since the characteristics of the host cell are generally not of interest at the endpoint of the experiment.
4. If the protocol you found for transforming lactobacillus is complicated, then it is undoubtedly because simpler protocols don't work.
(continued next comment)