Gene cloning

Cloning and expression analysis of cysteine protease gene (MwCP) in Agropyron mongolicum Keng

T. G. B. Y. Ao, Lang, M. L., Li, Y. Q., Zhao, Y., Wang, L. C., Yang, X. J., Ao, T. G. B. Y., Lang, M. L., Li, Y. Q., Zhao, Y., Wang, L. C., Yang, X. J., Ao, T. G. B. Y., Lang, M. L., Li, Y. Q., Zhao, Y., Wang, L. C., and Yang, X. J., Cloning and expression analysis of cysteine protease gene (MwCP) in Agropyron mongolicum Keng, vol. 15, p. -, 2016.

In this study, a cysteine protease gene (MwCP) from Agropyron mongolicum Keng was isolated using RACE. Sequence analysis indicated that MwCP was 1473 bp, and it contained a 1134-bp open reading frame, which encoded 377 amino acids with a 24-amino acid N-terminal signal peptide. The results indicated that the MwCP protein was a new member of the papain C1A family, and it was predicted to be an extracellular, secretory stable hydrophilic protein.

Cloning and characterization of up-regulated HbSINA4 gene induced by drought stress in Tibetan hulless barley

H. J. Yuan, Luo, X. M., Nyima, T. S., Wang, Y. L., Xu, Q. J., and Zeng, X. Q., Cloning and characterization of up-regulated HbSINA4 gene induced by drought stress in Tibetan hulless barley, vol. 14. pp. 15312-15319, 2015.

Hulless barley is an important crop cereal in Tibetan, China. Drought is a major abiotic stress in barley production. In this study, we cloned the drought-related HbSINA4 gene from the variety ‘Himalaya 10’ and analyzed its expression patterns under different drought and rehydration conditions. The cDNA of HbSINA4 was 1052 bp long, including an open reading frame of 771 bp that encoded a protein of 256 amino acids. The molecular weight of HbSINA4 protein was predicted to be 29.53 kDa and the theoretical pI was 8.32.

Cloning of superoxide dismutase from post-harvest Hami melon and quantitative expression analysis before and after disease

C. H. Shan, Tang, F. X., Chen, W., and Ma, W. R., Cloning of superoxide dismutase from post-harvest Hami melon and quantitative expression analysis before and after disease, vol. 14, pp. 18229-18240, 2015.

Primers were designed according to the Cu/Zn-SOD gene sequences of cloned Cucurbits plants (cucumbers and watermelons) available in NCBI. Total RNA from Hami melon pulp was used as a template. Following RT-PCR amplification, a 403-bp fragment of the Hami melon Cu/Zn-SOD gene was obtained. According to alignment in BLAST and phylogenetic tree analysis, the cloned gene fragment was confirmed to be the Hami melon Cu/Zn-SOD gene sequence.

Cloning and characterization of the nicotianamine synthase gene in Eruca vesicaria subsp sativa

B. L. Huang, Cheng, C., Zhang, G. Y., Su, J. J., Zhi, Y., Xu, S. S., Cai, D. T., Zhang, X. K., and Huang, B. Q., Cloning and characterization of the nicotianamine synthase gene in Eruca vesicaria subsp sativa, vol. 14, pp. 18121-18130, 2015.

Nicotianamine (NA) is a ubiquitous metabolite in plants that bind heavy metals, is crucial for metal homeostasis, and is also an important metal chelator that facilitates long-distance metal transport and sequestration. NA synthesis is catalyzed by the enzyme nicotianamine synthase (NAS). Eruca vesicaria subsp sativa is highly tolerant to Ni, Pb, and Zn. In this study, a gene encoding EvNAS was cloned and characterized in E. vesicaria subsp sativa.

Ectopic expression of the BoTFL1-like gene of Bambusa oldhamii delays blossoming in Arabidopsis thaliana and rescues the tfl1 mutant phenotype

H. Y. Zeng, Lu, Y. T., Yang, X. M., Xu, Y. H., and Lin, X. C., Ectopic expression of the BoTFL1-like gene of Bambusa oldhamii delays blossoming in Arabidopsis thaliana and rescues the tfl1 mutant phenotype, vol. 14, pp. 9306-9317, 2015.

TERMINAL FLOWER1 (TFL1) homologous genes play major roles in maintaining vegetative growth and inflorescence meristem characteristics in various plant species; however, to date, the function of the bamboo TFL1 homologous gene has not been described. In this study, a TFL1 homologous gene was isolated from Bambusa oldhamii and designated as BoTFL1-like. Phylogenetic analysis of TFL1 homologous genes revealed that BoTFL1-like shared more than 90% identity with the TFL1 genes of other Gramineae.

Cloning and transformation analysis of isoflavone synthase gene into Minshan Trifolium pratense

H. H. Hu, Jing, C. Q., Liu, R., Li, W. D., and Feng, H. G., Cloning and transformation analysis of isoflavone synthase gene into Minshan Trifolium pratense, vol. 14, pp. 9291-9297, 2015.

The aim of this study was to clone the isoflavone synthase (IFS) gene and establish the recombinant Minshan Trifolium pratense. The IFS gene was cloned from the callus of Minshan T. pratense using reverse transcription-polymerase chain reaction. The plant expression vector pRI101-AN-IFS was constructed and introduced into Agrobacterium tumefaciens strain LBA4404, and then screened under cephalosporin. IFS expression was detected by reverse transcription-polymerase chain reaction. The IFS gene was cloned successfully.

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