Molecular Cloning and Analysis of EuREF 1 Involved into Rubber Biosynthesis in Eucommia ulmoides Olive.
The cDNA,EuREF 1,encoding rubber elongation factor in Eucommia ulmoides Olive.was cloned using the method of RACE with the total mRNA of gutta-containing organization as the template.The RACE product1 075 bp fragment of EuREF 1 was ligated and cloned into the pGEM-T vector and sequenced.The results showed that the ORF ofEuREF 1 had 678 bp,which encodes a 225 amino acid of protein.The EuREF 1 sequence had 42％,41％ homology to the REF protein sequence of Vitis vinifera,Ricinus communis,respectively.The results ofblastp in GenBank suggested that EuREF 1 belongs to REF superfamily.The dicots expression vector,constructed by pSH-737 vector,pSH-35S-EuREF 1 was transformed into Nicotiana.tabacum L.mediated with Agrobaeterium tumefaeiens.The 43 transgenic tobacco lines were identified by PCR molecular detection and GUS histochemical staining.The transgenic tobacco exhibited obviously different in the leafphenotype compared with wildtype after four month.
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