Cloning Of The Downstream Sequence Of The Cotton Promoter Prom6 And The Functional Analysis Of The Gene GhF1 Specifically Expressed In Cotton Fibers

Abstracts In agricultural area, separating the functional gene possessing important agronomic traits, and performing molecular breeding through transgenic technology, which have important significance on modifying characteristics of crops varieties. Therefore, it is an excellent approach for genetic engineering schemes to modify cotton varieties that isolating and characterizing the genes correlating with fiber development as the objective gene of transforming cotton. In the study, according to the known pnm6 promoter sequence, we designed a pair of specific primer, and filtrated the cotton BAC through the way of mixing basin, in order to obtaining the BAC clone possessing the objective gene. We affirmed the BAC clone through southern blot hybridization, and subcloned the segment that hybrid with probe. After sequencing, we obtained 1kb new nucleotide sequence downstream of prom6. We further used the ligation-mediated PCR for chromosome walking and got 1.2kb new nucleotide sequence. Sequence analysis and homology comparison found that there has no gene in the 2.2 kb new sequence downstream of prom6.Arabidopsis is a model plant for studying plant genes, from the view of systems development, the mechanism controlling the cotton fibers developed from the outmost cells of the ovelus is similar to that of controlling the arabidopsis trichome development. Therefore, it has developed to be an excellent model system for studying the function of cotton genes resent years. In our study, we constructed plant expression vectors, GhFl that is specifically expressed in cotton fiber was respectively cloned into the vector PZP that driven by 35S and E6 promoter, and transformed the two plant expression vectors to arabidopsis. We filtrated transgenic arabidopsis, and performed PCR, then obtained the transgenic lines, morphological studies showed that transgenic plant have some phenotypic changes: the position of siliques were anomalistic; the siliques were significantly shorter and seed development was largely impaired; some plant were very short. Thus, it is possible that GhFl might play a role during the cotton fiber cell developmentFission yeast is simple single-celled biology, it has the advantages of easy to culture, reproducing fast, easy to gene manipulate, and having many similarities with advanced eukaryotic biology. We constructed GhFl into the S.pombe/E.coli shuttle vector pREP5N in sense(+) orientation, and transformed the resultant plasmid pREP5N+GhF1 to yeast cells. Morphological studies showed that the cells carrying pREP5N+GAF1 vector grown in induced media were much shorter compared to those harboring the same vector but grown in non-induction media, those yeast cells with pREP5N vector and untransformed yeast cells grown in induced media, undirected indicating that GhFl have function in the cotton fiber elongation.