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Cloning and Characterization of OsSKIPa gene from rice (Oryza sativa Linn. Var. KDML 105) |
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| รหัสดีโอไอ | |
| Creator | Suphawadee Ngorian;Pongsacorn Sanvittayakul;Paranee Sawangsri;Rungnapha Pitaktansakul;Phummarin Wanichananan;Hathairat Urairong |
| Title | Cloning and Characterization of OsSKIPa gene from rice (Oryza sativa Linn. Var. KDML 105) |
| Publisher | Department of Agriculture |
| Publication Year | 2559 |
| Journal Title | THAI AGRICULTURAL RESEARCH JOURNAL |
| Journal Vol. | 34 |
| Journal No. | 1 |
| Page no. | 13-27 |
| Keyword | OsSKIPa Gene, abiotic stress, cloning, characterization, plant expression vector, gene cassette |
| ISSN | 1258389 |
| Abstract | SKIPa gene plays role as critical and necessary gene affecting cell viability andenhances cell's ability to survive from abiotic stress such as drought and salinity. Thisresearch study aimed to clone OsSKIPa gene from KDML 105 rice and to inventexpression cassette for applying to plant variety improvement particularly against abioticstress by gene transformation technique. The research was done at the BiotechnologyResearch and Development Office, Department of Agriculture, Pathumthani Provinceduring May September 2011. In this study, full-length genomic DNA sequences of rice(Oryza Sativa Linn.) encoded to OsSKIPa was isolated from rice via PCR based method.The gene sequence contains a fragment of 2,353 bp, including a 1,824 bp complete ORF,the 5'UTR of 130 bp, 3'UTR of 399 bp and a polyadenylation signal ATAAA motif.OsSKIPa gene can be encoded to 607 amino acids polypeptide with molecular weight of81.2 kDa and has putative function as SKIP / SNW domain containing protein. The highlyconserved region of the gene is SNW/SKI-interacting protein-like, SNW/SKI-interactingprotein, transcript variant X1, transcript variant X2 and transcript variant X3 which arefound in Oryza sativa L. (NM001054719.1), Oryza brachyantha (XM006649000.1) andBrachypodium distachyon L. (XM010237787.2, XM010237788.2, XM010237789.2) with 99, 94 and 86% of homology, respectively. A1,824 bp fragment of OsSKIPa gene wasinserted into plant expression vectorpCAMBIA2300 containing 35SCaMV promoter and NOS terminator. NPTII was used as a selectable marker in this selection. It was found that the total size of derived over-expression cassette (pCAMBIA2300 OsSKIPa) was 11.5 kb. |
