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Updated: Sep 26, 2026
Functional Cloning Using a Xenopus Oocyte Expression System
Published on: January 30, 2016
Cloning and expression of ornithine decarboxylase gene from human colorectal carcinoma
Hai-Yan Hu1, Xian-Xi Liu, Chun-Ying Jiang
1Experimental Centre of Medical Molecular Biology, School of Medicine, Shandon University, Jinan 250012, Shandong Province, China.
Aim:
To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.
Methods:
Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing, the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.
Results:
ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E. coli M15 and expressed. The expressed ODC protein was verified with Western blotting.
Conclusion:
The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.
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