Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
Effect of lanthanum on expression of recombinant allophycocyanin gene in Pichia pastotis | |
Ren, YH; Ge, BS; Jin, HZ; Tang, ZH; Guo, CH; Qin, S | |
2005-08-01 | |
发表期刊 | JOURNAL OF RARE EARTHS
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ISSN | 1002-0721 |
卷号 | 23期号:4页码:491-495 |
文章类型 | Article |
摘要 | The enhancing effect of lanthanum on gene expression of recombinant allophycocyanin (rAPC), a potential antitumor medicine, in Pichia pastoris was studied. PCR and sequence analysis were used in order to prove whether the APC gene had integrated into the yeast genome. The expression level of the recombinant allophycocyanin (rAPC) in BMMY medium containing LaCl3 was detected by ELISA method. The recombinant allophycocyanin was determined by Western blot. The results show that the recombinant Pichia pastoris chromosome contained allophycocyanin gene. Expression efficiency of rAPC gene in Pichia pastoris was promoted by proper LaCl3 concentration like 2, 5, 10 mmol (.) L-1, among which 5 mmol (.) L-1 was the most effective. The highest expression yield of rAPC in the BMMY medium containing 5 mmol (.) L-1 LaCl3 was 4.4 mg (.) L-1 at 48 h, that was increased by 110% compared with 2.1 mg (.) L-1 of control, in the meantime, the optimum culture time is shortened from 72 to 48 h. The result of western blot analysis indicates that the rAPC consisted of two kinds of subunits with molecular weight of 19 and 21 kDa respectively.; The enhancing effect of lanthanum on gene expression of recombinant allophycocyanin (rAPC), a potential antitumor medicine, in Pichia pastoris was studied. PCR and sequence analysis were used in order to prove whether the APC gene had integrated into the yeast genome. The expression level of the recombinant allophycocyanin (rAPC) in BMMY medium containing LaCl3 was detected by ELISA method. The recombinant allophycocyanin was determined by Western blot. The results show that the recombinant Pichia pastoris chromosome contained allophycocyanin gene. Expression efficiency of rAPC gene in Pichia pastoris was promoted by proper LaCl3 concentration like 2, 5, 10 mmol (.) L-1, among which 5 mmol (.) L-1 was the most effective. The highest expression yield of rAPC in the BMMY medium containing 5 mmol (.) L-1 LaCl3 was 4.4 mg (.) L-1 at 48 h, that was increased by 110% compared with 2.1 mg (.) L-1 of control, in the meantime, the optimum culture time is shortened from 72 to 48 h. The result of western blot analysis indicates that the rAPC consisted of two kinds of subunits with molecular weight of 19 and 21 kDa respectively. |
关键词 | Lanthanum Chloride Recombinant Allophycoeyanin Expression Pichia Pastoris Rare Earths |
学科领域 | Chemistry, Applied |
收录类别 | SCI |
语种 | 英语 |
WOS记录号 | WOS:000232021000023 |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.qdio.ac.cn/handle/337002/3122 |
专题 | 实验海洋生物学重点实验室 |
作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Yantai Univ, Dept Biochem, Yantai 264005, Peoples R China 3.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China |
推荐引用方式 GB/T 7714 | Ren, YH,Ge, BS,Jin, HZ,et al. Effect of lanthanum on expression of recombinant allophycocyanin gene in Pichia pastotis[J]. JOURNAL OF RARE EARTHS,2005,23(4):491-495. |
APA | Ren, YH,Ge, BS,Jin, HZ,Tang, ZH,Guo, CH,&Qin, S.(2005).Effect of lanthanum on expression of recombinant allophycocyanin gene in Pichia pastotis.JOURNAL OF RARE EARTHS,23(4),491-495. |
MLA | Ren, YH,et al."Effect of lanthanum on expression of recombinant allophycocyanin gene in Pichia pastotis".JOURNAL OF RARE EARTHS 23.4(2005):491-495. |
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