Microbial DNA Technologies

This set of flashcards encompasses key concepts related to Microbial DNA Technologies, focusing on restriction enzymes, cloning vectors, PCR processes, and applications in biotechnology.

What are restriction enzymes and their primary function?

Restriction enzymes are proteins that recognize and cleave specific DNA sequences, producing either blunt or sticky ends.

What is the purpose of reverse transcriptase in biotechnology?

Reverse transcriptase synthesizes double-stranded DNA from RNA templates, allowing cDNA to be cloned without RNA processing.

List three essential features of a cloning vector.

A cloning vector must have an origin of replication, unique restriction sites, and a selectable marker.

Differentiate between plasmids and phage-based cloning vectors.

Plasmids are circular DNA molecules that replicate independently, while phage-based vectors are modified phage genomes that can insert DNA into host cells.

How is DNA inserted into a cloning vector in vitro?

DNA can be inserted into a cloning vector by cutting both with the same restriction enzyme to create compatible ends that can be ligated together.

What are the three steps of the PCR cycle and their functions?

The three steps are denaturation (separating DNA strands), annealing (binding primers to template), and extension (synthesizing new DNA strands).

What distinguishes real-time PCR from end-point PCR?

Real-time PCR quantifies DNA in a sample as it amplifies, while end-point PCR measures the quantity after amplification is complete.

What is the benefit of genomic and metagenomic libraries?

They provide a comprehensive repository of DNA from either a single organism or environment, allowing researchers to identify genes of interest.

Describe the role of expression vectors in biotechnology.

Expression vectors contain sequences that enable the transcription and translation of cloned genes, optimizing protein production in host cells.

What is Cas9 and its significance in gene editing?

Cas9 is an endonuclease used in CRISPR technology to cut DNA at specific sites, allowing modification of genomes.

What is directed evolution in microbial biotechnology?

Directed evolution involves making specific mutations in genes to enhance desired traits in production strains.

What is synthetic biology and its advantages?

Synthetic biology is the design of biological systems for novel functions, providing efficient assembly, reduced metabolic engineering, and combinatorial regulation.

Explain the process of gel electrophoresis in DNA analysis.

Gel electrophoresis separates DNA fragments by size, allowing visualization and analysis of the DNA after migration through an agarose gel.

What is the function of a His-tag in protein purification?

A His-tag on a protein facilitates its purification by using metal ion resins to capture proteins with high affinity for metal ions.

How does PCR enable the amplification of specific DNA sequences?

PCR uses specific primers complementary to target sequences to amplify only that DNA, despite the presence of competing sequences.