Recombinant DNA Technology Flashcards

Vocabulary flashcards based on lecture notes covering recombinant DNA technology, restriction endonucleases, vectors, and host cell transformation techniques.

Biotechnology

The science of using living organisms to produce beneficial products for humans, such as the use of yeast in bread making, selective breeding, or genetic engineering.

Genetic Engineering

The use of modern methods to manipulate genes or create new genes to produce organisms with desired characteristics.

Recombinant DNA Technology

The technique used in genetic engineering involving stages to isolate, insert, transform, screen, and express target genes.

Restriction Endonuclease

An enzyme produced in bacteria to cut viral DNA, which recognizes and cuts DNA at specific palindromic sequences.

Restriction Site

A specific, usually palindromic sequence of bases in DNA recognized and cut by a specific restriction endonuclease.

Sticky Ends

Staggered cuts with single-stranded ends produced on DNA fragments by certain restriction endonucleases, which can rejoin with matching fragments.

Blunt Ends

Straight cuts made directly across both strands of DNA by certain restriction endonucleases.

Complementary DNA (cDNA)

DNA synthesized from functional mRNA using reverse transcriptase, enabling bacteria to express eukaryotic genes without introns.

Reverse Transcriptase

An enzyme obtained from retroviruses that synthesizes complementary DNA (cDNA) from an mRNA template.

Eco RI

A restriction endonuclease enzyme originating from E. coli that cuts DNA at the specific sequence GAATTC / CTTAAG.

Bam HI

A restriction endonuclease enzyme originating from Bacillus amyloquefacein that cuts DNA at the specific sequence GGATCC / CCTAGG.

Pov II

A restriction endonuclease enzyme originating from Proteus vulgaris that cuts DNA at the specific sequence CAGCTG / GTCGAC.

Sma I

A restriction endonuclease enzyme originating from Serrana marcescens that cuts DNA at the specific sequence CCCGGG / GGGCCC.

Vector

A DNA vehicle that accepts foreign DNA at an MCS, replicates independently via an Ori, and contains a selectable marker.

Multiple Cloning Sites (MCS)

A specific region within a plasmid vector containing unique restriction sites that can be cut by specific restriction enzymes.

Origin of Replication Initiation (Ori)

A sequence in a vector plasmid that allows it to replicate independently of the host chromosome.

Selectable Marker

A gene on a vector (such as the Amp gene for ampicillin resistance) used to screen and identify host cells containing recombinant DNA.

Plasmid

A small circular double-stranded DNA found in bacteria (e.g., pUC18) carrying antibiotic resistance genes and capable of carrying $10\text{-}15\,\text{kbs}$ of inserted gene.

Bacteriophage

A virus containing DNA that infects bacteria (e.g., λ 2001), disabled for use as a vector capable of carrying up to $40\,\text{kbs}$ of inserted gene.

Cosmid

A plasmid vector constructed by inserting cos sequences from λ phage (e.g., sCOS-I), allowing it to carry up to $40\,\text{kbs}$ of inserted gene.

Yeast Artificial Chromosome (YAC)

An artificially constructed chromosome (e.g., pYAC) containing genes needed for replication in yeast cells, capable of carrying $100\text{-}3000\,\text{kbs}$ of inserted gene.

DNA Ligase

An enzyme used to join and seal sticky ends together between the target gene fragment and vector plasmid, creating recombinant DNA.

Host Cell

A cell (usually a bacterium) that receives recombinant DNA via transformation, maintains its structure across generations, and amplifies the gene product.

Transformation

The process of introducing new DNA or a recombinant plasmid into a host bacterial cell.

Calcium Chloride Heat Shock Method

A transformation method where bacteria are treated with calcium chloride solution and given a brief $42\,^\circ\text{C}$ heat shock to create temporary holes in the membrane for plasmid entry.

Gene Gun

A device used in transformation that shoots tiny DNA-coated metallic (gold or tungsten) particles into target living cells.

Clones

A population of genetically similar bacteria formed through the reproduction of a transformed host cell.