Cloning

Cloning and sequence analysis of an actin gene in aloe

S. S. Wen, He, D. W., Liao, C. M., Li, J., Wen, G. Q., and Liu, X. H., Cloning and sequence analysis of an actin gene in aloe, vol. 13, pp. 4949-4955, 2014.

Aloe (Aloe spp), containing abundant polysaccharides and numerous bioactive ingredients, has remarkable medical, ornamental, calleidic, and edible values. In the present study, the total RNA was extracted from aloe leaf tissue. The isolated high-quality RNA was further used to clone actin gene by using reverse transcription-polymerase chain reaction (RT-PCR). The result of sequence analysis for the amplified fragment revealed that the cloned actin gene was 1012 bp in length (GenBank accession No.

DNA barcoding for species identification in the Palmae family

A. Naeem, Khan, A. A., Cheema, H. M. N., Khan, I. A., and Buerkert, A., DNA barcoding for species identification in the Palmae family, vol. 13, pp. 10341-10348, 2014.

DNA barcoding is a promising tool for species identification at the molecular level. The barcoding system is well established for species differentiation in animals, while it is less common in plants. We evaluated 2 barcoding regions, maturase K (matK) and ribulose bisphosphate carboxylase (rbcL), to compare species of Palmae according to amplification success, discrimination power, and inter- and intra-specific divergence.

 Cloning and expression of UDP-glucose: flavonoid 3-O-glucosyltransferase gene in peach flowers

X. C. Wen, Han, J., Leng, X. P., Ma, R. J., Jiang, W. B., and Fang, J. G.,  Cloning and expression of UDP-glucose: flavonoid 3-O-glucosyltransferase gene in peach flowers, vol. 13, pp. 10067-10075, 2014.

To elucidate the connection between flower coloration and the expression of genes associated with anthocyanin biosynthesis, a gene encoding UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT) was isolated, and the expression of the last four genes in the anthocyanin biosynthetic pathway during peach flower development was determined. The nucleotide sequence of the peach UFGT (GenBank accession No. JX149550) is highly similar to its homologs in other plants.

Identification and bioinformatic analysis of a putative calcium-dependent protein kinase (CDPK6) from Toxoplasma gondii

N. Z. Zhang, Huang, S. Y., Zhou, D. H., Xu, Y., He, J. J., and Zhu, X. Q., Identification and bioinformatic analysis of a putative calcium-dependent protein kinase (CDPK6) from Toxoplasma gondii, vol. 13, pp. 10669-10677, 2014.

Toxoplasma gondii is recognized as an opportunistic human pathogen with a worldwide distribution. Development of effective vaccines is considered the only ideal way to control T. gondii infection. However, only one live vaccine is commercially available for use in sheep and goats. Therefore, the identification of more effective antigenic proteins is very important. In this study, we identified a novel putative calcium-dependent protein kinase of T.

Cloning, expression analysis and sequence prediction of the CCAAT/enhancer-binding protein alpha gene of Qinchuan cattle

H. Wang, Zan, L. S., Wang, H. B., Gong, C., and Fu, C. Z., Cloning, expression analysis and sequence prediction of the CCAAT/enhancer-binding protein alpha gene of Qinchuan cattle, vol. 11, pp. 1651-1661, 2012.

CCAAT/enhancer-binding protein alpha (C/EBPα) is an essential transcription factor, regulating the differentiation of adipocytes. We cloned the complete open reading frame of C/EBPα gene of Qinchuan cattle and analyzed its protein structures and expression profile in 15 tissues via DNA cloning, sequencing and RT-PCR. Analysis of the putative protein sequences revealed that C/EBPα consists of alpha helices, random coils and a few extended strands. A significant transmembrane structure was observed in amino acid region 233 to 252.

Construction and preliminary characterization of a river buffalo bacterial artificial chromosome library

N. B. Stafuzza, Abbey, C. A., Gill, C. A., Womack, J. E., and Amaral, M. E. J., Construction and preliminary characterization of a river buffalo bacterial artificial chromosome library, vol. 11, pp. 3013-3019, 2012.

River buffalo genome analyses have advanced significantly in the last decade, and the genome sequence of Bubalus bubalis will be available shortly. Nonetheless, large-insert DNA library resources such as bacterial artificial chromosomes (BAC) are still required for validation and accurate assembly of the genome sequence. We constructed a river buffalo BAC library containing 52,224 clones with an average insert size of 97 kb, representing 1.7 × coverage of the genome.

Molecular cloning, sequence characterization of a novel pepper gene NADP-ICDH and its effect on cytoplasmic male sterility

M. H. Deng, Wen, J. F., Huo, J. L., Zhu, H. S., Dai, X. Z., Zhang, Z. Q., Zhou, H., and Zou, X. X., Molecular cloning, sequence characterization of a novel pepper gene NADP-ICDH and its effect on cytoplasmic male sterility, vol. 11, pp. 3020-3031, 2012.

NADP-dependent isocitrate dehydrogenase (NADP-ICDH) is an important enzyme involved in energy metabolism. The complete coding sequence of the pepper (Capsicum annuum) NADP-ICDH gene was amplified using a reverse transcriptase PCR based on the conserved sequence information of the tomato and other Solanaceae plants and known highly homologous pepper ESTs.

Expression of splice variants of cancer-testis genes ODF3 and ODF4 in the testis of a prostate cancer patient

S. Ghafouri-Fard, Ghafouri-Fard, S., and Modarressi, M. H., Expression of splice variants of cancer-testis genes ODF3 and ODF4 in the testis of a prostate cancer patient, vol. 11. pp. 3642-3648, 2012.

The outer dense fiber (ODF) genes encode proteins that co-assemble along the axoneme of the sperm tail. Recently, it was demonstrated that some ODF genes are aberrantly expressed in tumors, including prostate adenocarcinoma, basal cell carcinoma, and chronic myeloid lymphoma. We cloned ODF3 and ODF4 cDNA from the testis of a patient suffering from prostate adenocarcinoma and found two alternative splice variants of these genes.

Molecular cloning and characterization of the pseudorabies virus US1 gene

M. L. Li, Chen, J. H., Zhao, Z. Y., Zhang, K. J., Li, Z., Li, J., Mai, J. Y., Zhu, X. M., and Cai, M. S., Molecular cloning and characterization of the pseudorabies virus US1 gene, vol. 12, pp. 85-98, 2013.

Using polymerase chain reaction, a 1050-bp sequence of the US1 gene was amplified from the pseudorabies virus (PRV) Becker strain genome; identification of the US1 gene was confirmed by further cloning and sequencing. Bioinformatics analysis indicated that the PRV US1 gene encodes a putative polypeptide with 349 amino acids.

Characterization of chicken natural resistance-associated macrophage protein encoding genes (Nramp1 and Nramp2) and association with salmonellosis resistance

X. M. He, Fang, M. X., Zhang, Z. T., Hu, Y. S., Jia, X. Z., He, D. L., Liang, S. D., Nie, Q. H., and Zhang, X. Q., Characterization of chicken natural resistance-associated macrophage protein encoding genes (Nramp1 and Nramp2) and association with salmonellosis resistance, vol. 12, pp. 618-630, 2013.

Natural resistance-associated macrophage protein 1 and 2 encoding genes (Nramp1 and Nramp2) are related to many diseases. We cloned the cDNA of chicken Nramp1 and Nramp2 genes, characterized their expression and polymorphisms, and investigated the association of some SNPs with resistance to salmonellosis. The Nramp1 cDNA was 1746 bp long and the Nramp2 cDNA was 1938 bp long. These cDNAs are similar to previously reported cDNAs, varying by two and one amino acids, respectively.

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