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#yeast artificial chromosome

3 public questions tagged with this topic.

Restriction Enzyme and Vector

Which feature makes YAC resemble natural yeast chromosomes?

Yeast artificial chromosomes were designed to replicate and segregate like native yeast chromosomes during mitosis and meiosis. Functionality requires three cis elements: autonomously replicating sequence acting as origin, centromere CEN conferring mitotic stability and ensuring proper spindle attachment and two telomeres protecting linear ends from degradation and fusion. URA3 and TRP1 provide selection, but structural resemblance relies on CEN, TEL and ARS combination. This tripartite core allows YAC to behave as an authentic linear chromosome, carrying megabase inserts and undergoing normal

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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Restriction Enzyme and Vector

Which is a selectable marker in yeast artificial chromosome (YAC)?

Yeast artificial chromosomes require selectable markers functional in auxotrophic yeast strains to ensure maintenance. URA3 encodes orotidine-5'-phosphate decarboxylase involved in uracil biosynthesis, complementing ura3 mutant phenotype. Yeast lacking functional URA3 cannot grow without uracil, so only transformants harboring YAC with URA3 survive on minimal medium. Similar markers include TRP1 for tryptophan and LEU2 for leucine selection. URA3 also allows counter-selection with 5-fluoroorotic acid for manipulating constructs. Bacterial markers like ampR are not selectable in yeast, making e

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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Restriction Enzyme and Vector

Which vector is best for cloning 200 kb DNA?

Cloning capacity is determined by vector structure and host tolerance for large inserts. Plasmids handle up to 10 kb, lambda phage 20 kb, cosmid around 45 kb, BAC 100-300 kb, YAC 100-2000 kb and mammalian artificial chromosome exceeds 2000 kb. For 200 kb genomic fragments, YAC is optimal among common options because it includes centromere, telomeres and ARS enabling segregation of very large linear eukaryotic DNA in yeast. BAC can accommodate up to 300 kb but 200 kb near upper limit with lower stability. YAC accommodates megabase inserts essential for genome mapping projects.

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

View Details