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produces growth hormone
o gene that codes for this protein is GH1 |
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infectious proteins that can cause degenerative brain disorders in mammals
• certain prion diseases are hereditary, but most become infected through directly ingesting prion proteins in food |
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responsible for a major exception to central dogma
• allows info to flow from RNA to DNA b/c catalyzes synthesis of DNA from RNA template |
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DNA that is produced from mRNA
• doesn’t have introns because mRNA doesn’t have introns |
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small, circular DNA molecule
• common in bacterial cells, but physically distinct from bacterial chromosome
• not required by cell for normal growth & reproduction
• most replicate independently of chromosome
• sometimes copied and transferred from one cell to another during conjugation
• can clone a gene by inserting it into a plasmid
• engineered plasmid would be replicated and passed onto daughter cells |
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| what a plasmid is called what it is used to carry DNA from another source |
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bacterial enzymes that cut DNA molecules at specific base sequences
• used to cut a gene out for later insertion into a cloning vector
• most only cut DNA at sites that form palindromes |
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o important b/c if restriction sites in different DNA sequences are cut with same restriction endonuclease, presence of sticky ends allows resulting fragments to be spliced together via complementary base pairing
• essence of recombinant DNA technology – ability to create novel combinations of DNA sequences by cutting specific sequences and pasting them into new locations |
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| marked copy of the molecule you’re looking for |
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single-stranded fragment of a labeled, known gene that binds to a single-stranded complementary sequence in the sample of DNA being analyzed
• by binding to target sequence, marks it
• must be labeled in some way so that it can be found after it has bound to the complementary sequence in the large sample of fragments |
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| how researchers can get millions/billions of copies of specific genes |
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| If a recombinant plasmid can be inserted into a bacterial or yeast cell, the foreign DNA will be copied and transmitted to new cells as the host cell grows and divides |
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| _____ often the vector of choice in genetic engineering, not ________ |
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| • cells that take up DNA from the environment and incorporate it into their genomes are said to undergo _______ |
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| to transform bacteria with a plasmid, researchers _________ |
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| increase permeability of cell’s plasma membranes using a specific chemical treatment or electric shock |
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| two options for cloning DNA |
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o inserting a gene into a bacterial plasmid
o polymerase chain reaction |
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| polymerase chain reaction (PCR) |
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an in vitro DNA synthesis reaction in which a specific section of DNA is replicated over and over to amplify the number of copies of that sequence
• technique for generating many identical copies of a particular section of DNA
• only possible when already have some info about DNA sequences near gene in question b/c have to start with short sequences of single-stranded DNA on either sides of gene in question |
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• denaturation, primer annealing and extension
• amount of template sequence doubles each time |
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o DNA polymerase found in bacterium Thermus aquaticus
• enzyme of choice in PCR b/c technique depends on heating reaction mix and is heat stable |
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| variation on basic in vitro DNA synthesis reaction to determine exact base sequences |
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| dideoxyribonucleoside triphosphates (ddNTPs) |
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identical to dNTPs found in DNA, except lack a hydroxyl group at 3’ carbon
• four types used in dideoxy sequencing
• ddATP, ddTTP, ddCTP, and ddGTP |
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| genetic map/linkage map/meiotic map |
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used to located gene(s) associated with a particular phenotype
• contains genetic markers |
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| records absolute position, in numbers of base pairs, along a chromosome |
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genes or other genetic loci or sites that have known locations
• each provides a landmark at a position along a chromosome that is known relative to other markers
• have to be polymorphic |
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| single nucleotide polymorphisms (SNPs) |
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site in DNA where some individuals in the population have different bases
• particularly abundant markers |
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| endonuclease recognition site |
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| short stretches of DNA where restriction endonucleases cut the double helix |
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| “across-genes,” have alleles that have been modified by genetic engineering |
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| discovery of Huntington's gene |
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• called newly discovered gene IT15 and its protein product huntingtin
• think that gradual buildup of huntingtin protein triggers neurons to undergo apoptosis |
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| in order for gene therapy to succeed, |
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• wild-type allele must be sequenced and understood
• must be a method for introducing a copy of the wild-type allele that ensures expression of the gene in the correct tissues, in the correct amount, at the correct time
o if defective allele is dominant, even more complication – might have to physically replace or block expression of existing dominant allele |
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o begins when a virus particle enters or attaches to host cell and inserts its genome into that host cell
• viruses that infect human cells can be used as vectors b/c sometimes becomes integrated into host-cell chromosome |
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o vector of choice in gene therapy
• viruses that have an RNA genome
• genomes include gene for reverse transcriptase, an enzyme that catalyzes production f double-stranded DNA from single-stranded RNA template |
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| serious problems with use of viruses as vectors |
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o usually cause disease
o retroviruses include HIV
o presence of viral proteins triggers immunal response that can cause dangerous side effects |
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| Ti (tumor-inducing) plasmid |
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contain several functionally distinct set of genes
• one set encodes products that allow bacterium to bind to cell walls of a host
• another set, called virulence genes, encodes proteins required to transfer part of the Ti DNA, called T-DNA (transferred DNA)
• offers an efficient way to introduce recombinant genes into plant cells |
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| tumorlike growth formed from a parasite |
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| independent of chromosome |
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