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The transgene must contain a promoter, which is a regulatory sequence that will determine where and when the transgene is active, an exon, a protein coding sequence (usually derived from the cDNA for the protein of interest), and a stop sequence.
Cre-Lox recombination involves the targeting of a specific sequence of DNA and splicing it with the help of an enzyme called Cre recombinase.Cre-Lox recombination is commonly used to circumvent embryonic lethality caused by systemic inactivation of many genes.
The vector itself generally carries a DNA sequence that consists of an insert (in this case the transgene) and a larger sequence that serves as the "backbone" of the vector. The purpose of a vector which transfers genetic information to another cell is typically to isolate, multiply, or express the insert in the target cell.
A DNA construct is an artificially-designed segment of DNA borne on a vector that can be used to incorporate genetic material into a target tissue or cell. [1] A DNA construct contains a DNA insert, called a transgene, delivered via a transformation vector which allows the insert sequence to be replicated and/or expressed in the target cell.
This 34 base pair (bp) loxP recognition site consists of two 13 bp palindromic sequences which flank an 8bp spacer region. The products of Cre-mediated recombination at loxP sites are dependent upon the location and relative orientation of the loxP sites. Two separate DNA species both containing loxP sites can undergo fusion as the result of ...
The T-DNA portion of the binary vector is flanked by left and right border sequences and may include a transgene as well as a plant selectable marker. Outside of the T-DNA, the binary vector also contains a bacterial selectable marker and an origin of replication (ori) for bacteria. [4] Representative series of binary vectors are listed below.
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If the transgene is incorporated into somatic cells, the transgene will stay with the somatic cell line, and thus its host organism. [ 6 ] Gene delivery is a necessary step in gene therapy for the introduction or silencing of a gene to promote a therapeutic outcome in patients and also has applications in the genetic modification of crops.