Modification of the n-terminal nucleotide sequence of mature hgm-csf results in high expression in the foreign host, anabaena sp strain pcc 7120
文献类型：期刊论文
| 作者 | Wei, Lanzhen; Ma, Weimin; Shi, Dingji; Wang, Quanxi |
| 刊名 | Journal of applied phycology
 |
| 出版日期 | 2006-04-01 |
| 卷号 | 18期号:2页码:153-159 |
| 关键词 | Anabaena sp strain pcc 7120 Gene modification Human granulocyte-macrophage colony stimulating factor Transgenic cyanobacterium Tri-parental conjugative transfer |
| ISSN号 | 0921-8971 |
| DOI | 10.1007/s10811-006-9086-y |
| 通讯作者 | Wang, quanxi(weilz@shnu.edu.cn) |
| 英文摘要 | Cloning and high foreign expression of the human granulocyte-macrophage colony stimulating factor (hgm-csf) gene were achieved in anabaena sp. strain pcc 7120 cells. to promote high expression of hgm-csf in cyanobacterial cells, pcr primers were designed to modify the n-terminal cdna sequence of mature hgm-csf, including a gc rich region and some discriminating against codons according to the degeneracy codon rules, selecting for prokaryotic usage codons. the pcr product encoding the modified hgm-csf was inserted downstream of the promoter, ppsba of the shuttle vector prl439, then ligated with pdc-08 to generate the shuttle expression plasmid, pdc-gm1. the resulting shuttle expression plasmid was transferred into the filamentous, heterocyst-forming cyanobacterium, anabaena sp. strain pcc 7120 using the tri-parental conjugation transfer method. the results of pcr amplification of wild type and transgenic cells indicated that the hgm-csf gene was successfully cloned into anabaena sp. strain pcc 7120 cells. western blot analysis showed that the protein expression of modified hgm-csf in transgenic cells harboring pdc-gm1 was 136% higher than that of non-modified hgm-csf in transgenic cells harboring pdc-gm0. additionally, there were similar rate of growth and content of chl a as compared to controls, suggesting that foreign hgm-csf did not impair the photosynthetic activity of host cells. taken together, the results indicate that modification of the n-terminal nucleotide sequence of mature hgm-csf results in high expression in the transgenic cells. |
| WOS关键词 | COLONY-STIMULATING FACTOR ; HIGH-LEVEL EXPRESSION ; HUMAN GM-CSF ; BIOLOGICAL-ACTIVITY ; ESCHERICHIA-COLI ; TRANSGENIC MICE ; ANIMAL-CELLS ; GENE ; CYANOBACTERIUM ; CLONING |
| WOS研究方向 | Biotechnology & Applied Microbiology ; Marine & Freshwater Biology |
| WOS类目 | Biotechnology & Applied Microbiology ; Marine & Freshwater Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000239891700006 |
| 出版者 | SPRINGER |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2379802 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Wang, Quanxi |
| 作者单位 | 1.Shanghai Normal Univ, Coll Life & Environm Sci, Shanghai 200234, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, Grad Sch, Shanghai 200032, Peoples R China 3.Chinese Acad Sci, Inst Bot, Beijing 100093, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wei, Lanzhen,Ma, Weimin,Shi, Dingji,et al. Modification of the n-terminal nucleotide sequence of mature hgm-csf results in high expression in the foreign host, anabaena sp strain pcc 7120[J]. Journal of applied phycology,2006,18(2):153-159. |
| APA | Wei, Lanzhen,Ma, Weimin,Shi, Dingji,&Wang, Quanxi.(2006).Modification of the n-terminal nucleotide sequence of mature hgm-csf results in high expression in the foreign host, anabaena sp strain pcc 7120.Journal of applied phycology,18(2),153-159. |
| MLA | Wei, Lanzhen,et al."Modification of the n-terminal nucleotide sequence of mature hgm-csf results in high expression in the foreign host, anabaena sp strain pcc 7120".Journal of applied phycology 18.2(2006):153-159. |
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来源：中国科学院大学
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