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Yeast artificial chromosome (YAC) vectors contain selectable markers. Loss of which marker at the cloning site distinguishes the re-ligated YACs from the original vector marker?
TRPI
SUP4
URA3
CEN
Answers (1)
The loss of the SUP4 marker at the cloning site distinguishes the re-ligated YACs from the original vector marker in yeast artificial chromosome (YAC) vectors.
The SUP4 marker is commonly used as a selectable marker in YAC vectors. It is derived from the SUP4 gene in yeast, which encodes a suppressor of nonsense mutations. When the foreign DNA fragment is inserted into the YAC vector during the cloning process, it disrupts the SUP4 marker.
The disruption of the SUP4 marker results in yeast cells being unable to suppress nonsense mutations, causing them to exhibit a specific phenotype, such as sensitivity to the antibiotic canavanine. This phenotype allows researchers to selectively grow yeast cells containing YAC vectors with intact SUP4 markers on media containing canavanine.
During the re-ligation process, if the foreign DNA fragment is lost or not properly re-inserted, the SUP4 marker at the cloning site can be restored. Consequently, yeast cells containing re-ligated YACs can regain the ability to suppress nonsense mutations and exhibit a different phenotype, such as canavanine resistance, distinguishing them from the original vector marker where the SUP4 marker is disrupted.
Hence, the correct answer is option 2.
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