|
Pak. J. Bot., 47(SI): 211-218, 2015. |
Back to Contents | ||||
|
|
Updated: 31-12-15 | ||||
|
|
CLONING AND AGROBACTERIUM MEDIATED TRANSFORMATION OF THE PUTATIVE PROMOTER REGION OF ORYZA SATIVA C3H52 GENE INTO LOCAL RICE VARIETY
Maleeha Khan1, Asad Jan1*, Saima Hashim1 and Khan Bahadar Marwat2
1The University of Agriculture, Peshawar-Pakistan 2SBBU, Upper Dir, Pakistan *Correspondence email: janasad@aup.edu.pk
Abstract: A number of zinc finger proteins are known to function in biotic and abiotic stress responses. The putative OsC3H52 gene promoter region, a member of CCCH zinc finger proteins family was isolated and characterized in this study. The in silico analysis of 1742bp promoter region of OsC3H52 gene using Plant care and Plant pan promoter analysis tools revealed the presence of ARE, TCA, G-BOX, TC rich repeats and MYBGAHV, GATABOX, WRKY71OS and Circadian cis-acting elements, respectively, which are involved in plant growth and development and stress responses. To clone the putative OsC3H52 gene promoter region, a promoter fragment of 1742 bp was amplified using indica rice (cv.Swat-1) genomic DNA. The putative OsC3H52 gene promoter region was cloned adjacent to GUS gene using pBIG binary vector and transformed into Agrobacterium tumefaciens (EHA105). For transformation of indica rice, cv. Swat-1, calli was induced from rice seed scutella using N6 medium supplemented with 2 mg/L 2,4-D and 0.5 mg/L kinetin. Six weeks old scutellar callli was used for Agrobacterium mediated transformation. The regenerated putative pBI POsC3H52: GUS transgenic plantlets were confirmed for transgene integration using PCR. Histochemical activity of the GUS gene in POsC3H52: GUS transgenic rice revealed its expression in calli and different parts of the plant including crown, leaf sheath, young leaves and emerging roots. The above results revealed that the putative OsC3H52 gene promoter is a functional promoter and it should be analyzed further in response to biotic and abiotic stress stimuli.
Key words: Indica rice, Zinc finger protein, Promoter, Cis-acting elements, GUS gene
|
||||
|
|
|||||
| Back to Contents |
|
Back to Contents | |||
|
|
|||||