Expression in vitro of OsPIN1a Gene and Analysis of Phosphorylation Sites
As one of PIN-FORMED(PIN) efflux carriers,OsPINla is involved in regulating auxin distributions in plant tissues.However,how these functional sites of OsPIN1a regulate hormone distribution still need to be further analyzed.In our study,OsPIN1a gene was isolated from Oryza sativa L.cv.Nipponbare using the specific primers,designed according to untranslated regions.To explore the phosphorylation sites of OsPIN1a in proteolytic structures,the expression vectors of single mutation[pET30-OsPIN1a(Ser233/Ala) and pET30-OsPIN1a(Ser254/Ala)]and double mutation[pET30-OsPIN1a(Ser233/Ala and Ser25,dAla)] were constructed by quickchange XL site-directed mutagenesis technique,respectively.OsPIN1a was expressed and analyzed by the expression system in vitro.Two phosphorylation sites of Ser-233 and Ser-254 can be phosphorylated by a serine-threonine protein kinase(PINOID,PID).The results provide a reference for further studying the mechanism of auxin polar transport.
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