Term
| hybrid DNA produced by joining the pieces of DNA from different organisms |
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Definition
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Term
| digests donor DNA and vector |
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Definition
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Term
| plasmid DNA or viral chromosome DNA that can carry a foreign DNA and transport the foreign DNA into a host cell |
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Definition
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Term
| most important DNA molecules that can be used as vectors and found in bacterium |
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Definition
| bacterial plasmids; bacteriophages |
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Term
| plasmids are small circular, double-strained molecule of DNA that can replicate independently of chromosomal DNA |
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Definition
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Term
| plasmids that can replicate by inserting themselves into the host chromosome |
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Definition
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Term
| carry only tra gene to conjugal transfer of plasmid |
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Definition
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Term
| carry genes that contribute to the host bacteria resistance to the antibacterial agents |
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Definition
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Term
| code for colicins, proteins that kill other bacteria |
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Definition
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Term
| allow the host bacterium to degradate unusual molecules |
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Definition
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Term
| confer pathogenicity on the host bacterium. Such as the Ti plasmids |
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Definition
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Term
| phages are viruses that specifically infect bacteria |
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Definition
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Term
| The genome DNA of lambda phage |
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Definition
| double stranded DNA, 49 kb in length |
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Term
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Definition
| can be in linear or circular form |
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Term
| complementery ends of linear form of lambda DNA |
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Definition
| sticky ends or cohesive ends |
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Term
| single-stranded DNA, length is 6407 nts; once inside the host cell it acts as template for synthesis of a complementery strand to form double-stranded DNA which is called replicative-form (RF) |
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Definition
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Term
| does not integrate into the host genome, but the phage particle assembles and releases continuously. The infected host can continue to grow and divide. |
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Definition
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Term
| Important for DNA manipulation and rDNA construction |
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Definition
| Nucelases, Ligases, Polymerases, DNA modifying enzymes |
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Term
| these are the enzymes that degrade DNA molecules by breaking the phosphodiester bonds between nucleotides in a DNA molecule |
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Definition
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Term
| catalyze the formation of a phosphodiester bond between 5' phosphate and 3' hydroxyl termini in DNA molecules |
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Definition
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Term
| enzymes that synthesize a new strand of DNA complimentary to an existing DNA or RNA template |
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Definition
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Term
| these enzymes modify DNA molecules by removal or addition of specific chemical groups |
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Definition
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Term
| cleave nucleotides from the ends of DNA molecules |
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Definition
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Term
| break the DNA molecules from the internal phosphodiester bonds within the DNA |
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Definition
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Term
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Definition
| repair single stranded breaks, or join together two double stranded DNA |
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Term
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Definition
| enzymes that synthesize a new strand of DNA complementary to an existing DNA or RNA template |
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Term
| To synthesize a new DNA strand, DNA polymerases request: |
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Definition
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Term
| short single stranded DNA complementary to template DNA or RNA. |
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Definition
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Term
| single stranded DNA or RNA used by DNA polymerase to synthesize new complementary DNA strand |
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Definition
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Term
| this enzyme has two functions DNA synthesis( polymerase) and DNA degradation (nuclease) |
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Definition
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Term
| the first 323 amino acids of the polypeptide (in DNA polymerase I) |
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Definition
| control the nuclease function of the enzyme (the rest have the polymerase function) |
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Term
| removing the first 323 amino acids of DNA polymerase I, the rest of the fragment retains polymerase function, but lose the nuclease activity |
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Definition
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Term
| specific DNA polymerase I which is heat-resistant and can function at high temperature; is a very important enzyme used in PCR |
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Definition
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Term
| DNA polymerase uses RNA as template to synthesize complementary DNA (cDNA) |
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Definition
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Term
| an enzyme that catalyzes the removal of 5' phosphate group from a DNA or RNA molecule |
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Definition
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Term
| an enzyme that catalyzes the transfer of a phosphate from ATP to the 5' end of a DNA or RNA molecule |
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Definition
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Term
| an enzyme that adds one or more deoxyribonucleotides onto the 3' terminus of a DNA molecule |
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Definition
| Terminal deoxynucleotidyl transferase |
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Term
| ideal enzymes to cut vector and donor DNA |
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Definition
| Restriction Endonucleases (RE) |
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Term
| enzymes isolated from bacteria that recognize specific sequence in DNA molecules and cleave DNA into fragments with specific ends: sticky ends or blunt ends. |
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Definition
| Restriction endonucleases |
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Term
| the most useful RE used in rDNA construction. Majority recognize specific DNA sequence 6 nucleotides in length. Some recognize 4, 5, or even longer nucleotide sequences. |
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Definition
| Type II Restriction Endonucleases |
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Term
| some REs recognize specific DNA sequence and cut the DNA into fragments with double strand ends |
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Definition
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Term
| REs recognize specific DNA sequence and cut DNA into fragments with short single-overhangs at each end |
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Definition
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Term
| have low efficiency of ligation. The ligation efficiency can be improved by increasing the concentration of DNA |
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Definition
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Term
| increase the efficiency of ligation. The complementary sticky ends can pair each other by hydrogen bonds to form double strands, which help ligase easily form phosphodiester bonds to increase the ligation efficiency |
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Definition
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Term
| Methods that are used to add sticky ends onto a DNA fragment |
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Definition
| DNA linkers, Adaptors, Producing sticky ends by adding homopolymer tail |
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Term
| are synthesized short dsDNA, the sequence is known. They contain one restriction sit and are blunt-ended |
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Definition
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Term
| synthesized short dsDNA with a sticky end. To avoid the ligation between them the 5' phosphate group of the sticky end is removed. After linkage to the blunt-ended DNA fragment the 5' phosphate group is added by polynucleotide kinase |
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Definition
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Term
| Using terminal transferase to add poly(dC) to the 3'-OH termini of blunt-ended vector DNA --> to be cloned |
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Definition
| Producing sticky ends by adding homopolymer tail |
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Term
| bacterial cells are treated by chemicals to enhance their ability to take up exogenous DNA |
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Definition
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Term
| Mix plasmid DNA with competent cells and incubate on ice for 30 min, then 42 Celsius for 30 sec-2 min |
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Definition
| Procedure of Transformation |
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Term
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Definition
| have ampicillin and tetracycline resistance gene |
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Term
| Identification of the recombinants wit the correct target gene insert |
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Definition
| Complementary nucleic acid strands hybridization-- southern blot. Use specific primers to amplify the target gene insert ---PCR. DNA sequence analysis |
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