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Molecular Biotechnology

by: Austen Pollich

Molecular Biotechnology CHM 53800

Marketplace > Purdue University > Chemistry > CHM 53800 > Molecular Biotechnology
Austen Pollich
GPA 3.75

Philip Low

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Philip Low
Class Notes
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This 3 page Class Notes was uploaded by Austen Pollich on Saturday September 19, 2015. The Class Notes belongs to CHM 53800 at Purdue University taught by Philip Low in Fall. Since its upload, it has received 66 views. For similar materials see /class/207969/chm-53800-purdue-university in Chemistry at Purdue University.


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Date Created: 09/19/15
MIDTERM EXAM VOCABULARY Annealing DNA or RNA to pair by hydrogen bonds to a complementary sequence forming a double stranded polynucleotide The term is often used to describe the binding of a DNA probe or the binding of a primer to a DNA strand during a polymerase chain reaction PCR The term is also often used to describe the reformation renaturation of complementary strands that were separated by heat thermally denatured Antisense the sequence of chromosomal DNA that is transcribed The DNA sequence complementary to all or part of a functional RNA An RNA sequence that is complementary to all or part of a functional RNA Autonomous replicating sequence any cloned DNA sequence that initiates and supports extrachromosomal replication ofa DNA molecule in a host cell yeast Autoradiography a technique that captures the image formed in a photographic emulsion as a result of the emission of either light or radioactivity from a labeled component that is placed next to an unexposed film Auxotroph a mutant that is unable to synthesize an essential amino acid and therefore the amino acid must be provided as a nutritional supplement for growth Bacterial artificial chromosome a vector system based on the EColi F factor plasmid that is used for cloning large 100300kb DNA inserts BLAST basic local alignment search tool a computer program for determining a match between a query sequence and a sequence in the database BLASTn compares a DNA query sequence to a DNA database BLASTp compares a DNA amino acid sequence to a protein database C39 quot 39 site a 39 u 39 location where foreign DNA can be integrated often without impacting any essential function in the host organism DNA hybridization the pairing of two DNA molecules often from different sources by hydrogen bonding between complementary nucleotides This technique is used to detect the presence of a specific nucleotide sequence in a DNA sample Expressed sequence tag a partially sequenced cDNA clone for which a PCR assay exists Fusion protein the product of two or more coding sequences from different genes that have been cloned together and that after translation form a single polypeptide sequence IMPACT ntein Mediated Purification with an Affinity Chitinbinding Tag utilizes the inducible self cleavage activity of engineered protein splicing elements termed inteins to purify recombinant proteins by a single affinity column This system distinguishes itself from other protein fusion systems by its ability to separate a recombinant protein from the affinity tag without the use of a protease n the presence of thiols such as DTT Bmercaptoethanol or cysteine the intein undergoes specific self cleavage which releases the target protein from the chitinbound intein tag IPTG sopropylBDthiogalacostopyranoside an inducer of the lac operon used to induce cloned genes that are under the control of the lac repressorlac promoter system Klenow fragment a product of proteolytic digestion of the DNA polymerase I from EColi that retains both polymerase and 3 exonuclease activity but not 5 exonuclease activity Marker peptide a portion of a fusion protein that facilitates its identification or purification MALDI MS mass spectrometry technique often used to determine the masses of peptides The peptides are mixed with a matrix consisting of an organic acid and then ionized with a laser The ions are accelerated through a tiny tube by using a highvoltage current and the time required for them to reach the ion detector is used to determine their molecular mass lower mass ions reach the detector first Northern blotting RNA that has been separated by PAGE is transferred to a nitrocellulose matrix and the presence of a specific RNA molecule is detected by DNARNA hybridization Palindromic sequence complementary DNA sequences that are the same when each strand is read in the same direction These types of sequences serve as recognition sites for type II restriction endonucleases Polylinker a synthetic DNA sequence that contains a number of different restriction endonuclease sites MCS Polymerase chain reaction a technique for amplifying a specific segment of DNA by using a thermostable DNA polymerase dNTPs and primer sequences in multiple cycles of denaturation renaturation and DNA synthesis Signal recognition complex a group of proteins that bind to the signal peptide of a newly synthesized protein and target the protein for secretion across the cytoplasmic membrane through the secretion complex Southern blotting a technique for transferring denatured DNA molecules that have been separated by PAGE electrophoresis to a nitrocellulose matrix on which a hybridization assay can be performed Stringency Hybridization conditions higher the stringency lower the probability of hybridization Increase the temperature or decreasing the salt concentration raises the stringency Targeting vector a cloning vector carrying a DNA sequence capable of participating in a crossover event at a specified chromosomal location in the host cell TATA box the DNA sequence to which RNA polymerase binds and that lies upstream from the site of initiation of transcription and ensures that transcription starts at a specified nucleotide also called Prinow Box in prokaryotes and Hogness box in eukaryotes Western blotting transfer of a protein from a gel to a nitrocellulose matri Yeast artificial chromosome a yeastbased vector system for cloning large gt100kb DNA inserts YAC


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