Cohesin complexes hold sister chromatids together whenever nuclear division occurs. It is important that the Cohesin complex is loaded onto the sister chromatids at the proper time and removed at the proper time. Two proteins involved in an antiestablishment complex that removes Cohesin from chromosomes are WAPL and PDS5. PDS5 also assists in Cohesin maintenance. Deficiency in WAPL has been shown to cause improper sister chromatid segregation, which can lead to diseases such as cancer. PDS5 deficiency can lead to improper maintenance of Cohesin. Arabidopsis has been shown to have two copies of WAPL and five copies of PDS5. Arabidopsis WAPL-1 cDNA has been cloned into pet22b vector for future overexpression, antibody production, and immunolocalization. WAPL-2 has been cloned into an RNAi2 vector with a meiosis specific promoter and into Agrobacterium, from where it will be transformed into Arabidopsis double homozygous Wapl mutants and the effects of Wapl depletion observed. PDS5-2 has been cloned into an RNAi1 vector with a meiosis specific promoter, from where it can be cloned into an RNAi2 vector and transformed into Arabidopsis to observe the effects of PDS5 depletion in meiotic cells.

Cloning of cohesin proteins for antibody production and RNA interference

247th National Meeting (2014)

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