14/ Shout-out to Zhihao Lai who developed this awesome vector-building system as a SIDE PROJECT during his PhD! 🧪 #Cloning#CloningSystems#plasmids#StructuralBiology#protein#sciencebsky.app/profile/davi...

CDdavidovichlab.bsky.socialOct 11, 2024 1:17pm

Have you always wanted to build multi-gene baculovirus expression vectors without the pain? We introduce MoClo Baculo: + No PCR + No primers + Swap tags/ORFs easily + No more sequencing gazillion intermediate plasmids + Bonus: yeast compatibility 🧵 www.biorxiv.org/content/10.1...

Modular cloning of multigene vectors for the baculovirus system and yeast

Recombinant macromolecular complexes are often produced by the baculovirus system, using multigene expression vectors. Yet, the construction of baculovirus-compatible multigene expression vectors is c...

12/ Fun fact #3: Intermediate plasmids (Level 1) can be used for protein expression, making MoClo Baculo useful even if you wish to quickly build vectors that express only a single protein. (Though, in such case, you'd like to sequence these plasmids).

11/ Fun fact #2: No sequencing of intermediate products! No PCR/primers are used --> low mutation rate --> intermediate plasmids ("Level 1") can be validated only by restriction digestion --> a whole plasmid sequencing can typically be done only on the final multi-gene plasmid.

10/ Fun fact #1: MoClo Baculo is fully compatible with the BacToBac system. If you use pFastBac1 plasmids or their derivatives, you can use your normal protein expression workflow. www.thermofisher.com/order/catalo...

9/ What "special reagents" do you need for using MoClo Baculo? Nothing special: + MoClo Baculo toolkit plasmids (soon will be on AddGene) + Some MoClo Yeast toolkit plasmids (on @addgene.bsky.social ) + Your ORFs without BsaI and BsmBI sites (for yeast compatibility, also without NotI).

7/ Does it work? Yes! As a demo, we constructed multi-gene baculovirus expression vectors of the 4- and 5-subunit PRC2 complexes (plasmids size: ~20 kbp).

Have you always wanted to build multi-gene baculovirus expression vectors without the pain? We introduce MoClo Baculo: + No PCR + No primers + Swap tags/ORFs easily + No more sequencing gazillion intermediate plasmids + Bonus: yeast compatibility 🧵 www.biorxiv.org/content/10.1...

Modular cloning of multigene vectors for the baculovirus system and yeast

Recombinant macromolecular complexes are often produced by the baculovirus system, using multigene expression vectors. Yet, the construction of baculovirus-compatible multigene expression vectors is c...

This phrasing makes it sound like he’s interested in inventing plasmids that let you throw balls of fire at people in an underwater city, and not just a really pseudo-intellectual form of racism

NMnick-martin.bsky.socialOct 10, 2024 3:43am

The New York Times just described Donald Trump's use of debunked and racist pseudoscience, which is at the heart of his perverted promises to purify the blood of the nation, merely as a "long-held fascination with genes and genetics." I'm floored.

Scientists didn't invent plasmids — nature did! Check out our recent blog post to learn about the natural plasmids all around you, and how they survive outside the lab. blog.addgene.org/pla...

Plasmids 101: The Wide World of Natural Plasmids

Where did plasmids come from? Learn more about natural plasmids, the benefits they offer cells, and how scientists have adapted them into laboratory tools.

Just out in PLOS Genetics: Surface exclusion of IncC conjugative plasmids and their relatives. journals.plos.org/plosgenetics...#plasmids#plasmidbiology

Surface exclusion of IncC conjugative plasmids and their relatives

Author summary Bacterial conjugation plays a pivotal role in the evolution of bacterial populations. The circulation of drug resistance genes bolsters the emergence of multidrug-resistant pathogens, w...