a. the dog CDNA coding for fucosidase b. a degenerate probe derived from the reported 20 amino acid sequence for human fucosidase c. the antibody to human fucosidase d. two of the above e. all of the above
I recently isolated the human enzyme called fucosidase and prepared an
antibody to it. Now I want to isolate a cDNA clone coding for this enzyme from a human cDNA
library. A friend of mine in the lab next door has informed me that he had recently isolated a
cDNA coding for dog fucosidase that I can use if I desire. In addition, I just read an article which
reported the sequence of the first 20 amino acids of human fucosidase. Which of the probes
listed below do you think I could use to screen my library to identify the cDNA clone containing
human fucosidase?
A probe is one of the essential components necessary to identify a gene during cloning. A probe is often a cloned bit of DNA that contains a fragment of the sequence that you are looking for. Typically, the probe is made radioactive and then immersed in a solution. The solution is then passed through filters containing immobilised clones. The idea here is that the probe will connect to any clone that contains sequences comparable to those present on the probe. This process of binding is known as hybridization.
Probe - a nucleic acid (typically DNA) that is complementary to a certain gene or nucleic acid sequence of interest; while screening a cDNA library, an antibody can be used to detect the protein encoded by the clone's insert.
Homologous Probe - a probe that is perfectly complementary to the nucleic acid sequence being searched; for example, a human cDNA used to search a human genomic library.
Heterologous Probe - a probe that is similar to, but not exactly complementary to, the nucleic acid sequence being searched; for example, a dog probe used to search a human genome library.
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