Construction of recombinant plasmid of chicken calbindin-D28k
To construct the eukaryotic expression vector of pCEP4-CaBP-D28k, a pair of primers was designed according to chicken calbindin-D28k (CaBP-D28k) gene sequence published in GenBank. Chicken CaB P-D28k gene was amplified by RT-PCR from chicken intestine total RNA extracts and was cloned into eukaryoti...
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| Language: | English |
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Zhejiang University Press
2008-11-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.06.004 |
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| author | YU Lu WANG Ya-qian ZHANG Shi-yuan LI Yan-wu LI Zhi-xue |
| author_facet | YU Lu WANG Ya-qian ZHANG Shi-yuan LI Yan-wu LI Zhi-xue |
| author_sort | YU Lu |
| collection | DOAJ |
| description | To construct the eukaryotic expression vector of pCEP4-CaBP-D28k, a pair of primers was designed according to chicken calbindin-D28k (CaBP-D28k) gene sequence published in GenBank. Chicken CaB P-D28k gene was amplified by RT-PCR from chicken intestine total RNA extracts and was cloned into eukaryotic expression vector pCEP4, and transferred into COS-1, The gene expression was detected by indirect immunofluorescent assay (IFA). The result shows that the cloned CaBP-D28k gene, encoding 261 amino acids, was 789 bp in length which was 99.9% identity but only one nucleotide changed from G to T at site 760 compared with published sequence from GenBank database. The homology of CaBP-D28k gene sequence between chicken and other species such as frog, house mouse, human was 79.7%, 78.2%, 78.6%, respectively; the target segment in recombinant plasmid was characterized and confirmed by restrictive endonuclease assay as well as sequence analysis. The expression of CaB P-D28k could be detected in COS-1. This research provides a novel means to study regulation mechanism of calcium metabolism in chickens through pCEP4-CaBP-D28k. |
| format | Article |
| id | doaj-art-6ef266f87d06481196e37c91d9ab3cf8 |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2008-11-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-6ef266f87d06481196e37c91d9ab3cf82025-08-20T03:58:14ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552008-11-013460861410.3785/j.issn.1008-9209.2008.06.00410089209Construction of recombinant plasmid of chicken calbindin-D28kYU LuWANG Ya-qianZHANG Shi-yuanLI Yan-wuLI Zhi-xueTo construct the eukaryotic expression vector of pCEP4-CaBP-D28k, a pair of primers was designed according to chicken calbindin-D28k (CaBP-D28k) gene sequence published in GenBank. Chicken CaB P-D28k gene was amplified by RT-PCR from chicken intestine total RNA extracts and was cloned into eukaryotic expression vector pCEP4, and transferred into COS-1, The gene expression was detected by indirect immunofluorescent assay (IFA). The result shows that the cloned CaBP-D28k gene, encoding 261 amino acids, was 789 bp in length which was 99.9% identity but only one nucleotide changed from G to T at site 760 compared with published sequence from GenBank database. The homology of CaBP-D28k gene sequence between chicken and other species such as frog, house mouse, human was 79.7%, 78.2%, 78.6%, respectively; the target segment in recombinant plasmid was characterized and confirmed by restrictive endonuclease assay as well as sequence analysis. The expression of CaB P-D28k could be detected in COS-1. This research provides a novel means to study regulation mechanism of calcium metabolism in chickens through pCEP4-CaBP-D28k.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.06.004chicken<italic>calbindin-D28k</italic> genegene cloningeukaryotic expression vectorindirect immunofluorescent assay (IFA) |
| spellingShingle | YU Lu WANG Ya-qian ZHANG Shi-yuan LI Yan-wu LI Zhi-xue Construction of recombinant plasmid of chicken calbindin-D28k 浙江大学学报. 农业与生命科学版 chicken <italic>calbindin-D28k</italic> gene gene cloning eukaryotic expression vector indirect immunofluorescent assay (IFA) |
| title | Construction of recombinant plasmid of chicken calbindin-D28k |
| title_full | Construction of recombinant plasmid of chicken calbindin-D28k |
| title_fullStr | Construction of recombinant plasmid of chicken calbindin-D28k |
| title_full_unstemmed | Construction of recombinant plasmid of chicken calbindin-D28k |
| title_short | Construction of recombinant plasmid of chicken calbindin-D28k |
| title_sort | construction of recombinant plasmid of chicken calbindin d28k |
| topic | chicken <italic>calbindin-D28k</italic> gene gene cloning eukaryotic expression vector indirect immunofluorescent assay (IFA) |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2008.06.004 |
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