BIOC0005 - SECTION 3 BACTERIAL GENOMICS EXAM QUESTIONS AND ANSWERS WITH COMPLETE SOLUTIONS VERIFIED LATEST UPDATE

BIOC0005 - SECTION 3 BACTERIAL GENOMICS EXAM

QUESTIONS AND ANSWERS WITH COMPLETE

SOLUTIONS VERIFIED LATEST UPDATE


List out the 5 main uses of plasmids in the lab

1. Gene cloning (cloning vector, expression vector)



2. Gene editing/engineering

(deliver CRISPR-Cas, gene knockout or knock in)



3. Reporter assay (reporter plasmids, fluorescent protein fusion)



4. Protein production and purification (expression plasmids, affinity tag plasmids)



5. Functional (gene overexpression, RNAi)

What are the 4 main considerations when using plasmids

1. PCN and size

2. Selection markers (antibiotic or other for screening)

3. Compatibility

4. Quality control (verify plasmid integrity, accuracy)

What 2 ways is replication initiation mediated by

, 1. Iteron-Mediated replication initiation

2. asRNA-Mediated replication initiation

Explain Iteron-Mediated replication initiation

Iteron = short DNA sq repeated w/i oriV



1. Iterons are recognition sites for replicase



2. Replicase binds and assembles replication and triggers unwinding

Explain asRNA mediated

asRNA = complementary to sense RNA transcripts



1. Binds to RNA and blocks protein binding



2. Regulates plasmid replication in response to environmental cue

What does plasmid incompatibility mean and reasons

Inability of plasmids to coexist and be maintained stably



Reasons: Same replication factors -> leads to competition

True or False? Ultra low copy number plasmids use plasmid partitioning while

higher PCN uses random distribution

True



Ultra low uses two methods