Identification of actin-related gene family from G. barbadense Giza 88 cultivar using PCR-based positional cloning [electronic resource].

Includes references.

Plant actins contribute strongly in cell cytoskeleton, microtubule filaments regulation and cellulose deposition orientation during cotton fiber cell development, which directly affects fiber quality. Identification of actin-related gene family from the Egyptian cotton is considered a corner stone in future engineering of fiber cell traits. P1-derived artificial chromosome (PAC) library has been constructed for the Egyptian extra long stable variety Giza88. The Giza88-PAC library comprised 8900 PAC clones with 70 Kb average size; representing 0.3 equivalents to the haploid genome (2118 Mb) of Gossypium barbadense. Randomly selected PAC clones were subjected to actin PCR-based screening using GhACT2 degenerate primers, which resulted in 14 actin positive clones. MPAC94 as one of these positives was purified and subjected to physical mapping and PCR-based positional cloning. The results indicated the recovery of a positive MPAC94/EcoRI actin fragment (16.26 Kb), which was confirmed by RT-PCR and sequence alignment at upland cotton database. This study is the first from its kind to identify one gene fragment of the actinrelated gene family in Egyptian cotton Giza88 using PCR-based positional cloning technology.

Summary in Arabic.

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