Yeast Plasmid Vectors, DNA Segments, Eukaryotic System, Plasmids Phages, Synthesize Leucine, Integrative Transformation

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Cloning and Expression Vectors Cloning and Expression Vectors Introduction Cloning Vectors for Recombinant DNA Plasmids as Vectors Three Phases of the Development of Plasmid Cloning Vectors Alpha-Complementation for Screening Using "blue-white"Clonies Nonsense Suppressors to Complement Termination Codons in Vectors E.Coli Plasmid Vectors pBR322 and pBr327 Vectors pUC Vectors pSP64-pSP65 pGEM-3, pGEM-4, pGEM-3Z, pGEM-4Z ΠAN13 from ΠAN7 Yeast Plasmid Vectors Retriever Vectors Agrobacterium Plasmid Vectors Lambda Phage (λ)Vectors Lambda Phage (Lysis and Lysogeny) Genetic Markers for the Selection and Screening of λ Vectors Development of Lambda (λ)Phage Vectors Insertion λ Vectors Replacement λ Vectors M13 Phage as a Vector for DNA Sequencing M13-vectors-of-mp-Series Hosts for M13 Vectors Cosmids as Vectors Phagemids as Vectors Phagemids pUC118 and pUC119 Phagemid pBluesript IIKS

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Yeast plasmid vectors

Special vectors are also available for introducing DNA segments in yeast cells, a eukaryotic system that has been used extensively for developing genetically engineered yeast cells. Although E. coli plasmids or phages can be used for transfer of genes to yeast cells, the frequency of transformation is rather low.

For instance, yeast cells that are LEU^- (lacking the ability to synthesize leucine) could be transformed using chimeric plasmid carrying LEU⁺ gene from yeast. This involved integrative transformation (crossing over) having low frequency. To overcome this difficulty of low frequency transformation, efforts were made' to prepare a variety of vectors for yeast.

The possible examples of the yeast plasmid vectors are:
(i) YIp or yeast integrative plasmids, which allow transformation by crossing over and have no replication origin;

(ii) YEp or yeast episomal plasmids, which carry 2µ DNA sequence influencing the origin of replication and rep genes;

(iii) YRp or yeast replicating plasmids, which carry autonomously replicating sequence; this sequence is very common with many yeast genes so that stable transformation can be achieved by crossing over;

(iv) YCp or yeast centromere (CEN) containing plasmids, which function as true chromosomes and segregate during mitosis and meiosis; (v) pYAC or yeast artificial chromosome, which carries both centromere and telomere sequences and, therefore, can be used to obtain artificial chromosomes.

Five different yeast plasmid vectors

A. no origin of replication; stable if integrate into yeast chromosome	B. 2m contains origin of replication and rep genes; high copy number; unstable	C. ARS allows autonomous replication; unstable; high copy number	D. contains centromere stable; single copy	E. contains centromere and telomeres; stable

The above vectors have been extensively utilized for a study of yeast genome and are also used as shuttle vectors, which allow genic sequences to be routinely transferred back and forth between yeast and E. coli cells, provided they contain origins of replication that are active in both yeast and E. coli.

These vectors also have markers enabling the selection of E. coli cells or colonies transformed with these vectors. The vectors can, therefore, be amplified in E. coli and then used to transform yeast cells.

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