Construction of Human Endostatin in Yeast Eukaryotic Expression Vector_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

ZHOU Dongfeng ¹ , ZHANG Zhongguang ²

Department of General Surgery, Qingdao Municipal Hospital, Qingdao 266011, China;

Corresponding author：

Keywords：

Human endostatin; Clone; Construction; Yeast

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Objective 　To construct yeast eukaryotic expression vector carrying human endostatin (ES) cDNA.　
Methods 　The functional fragment of endostatin gene in human hepatic tissue was amplified by using RT-PCR technology, and cloned into yeast pPIC9 expression vector. The positive clone was sequenced by using automatized sequencer.
Results 　The endostatin cDNA was successfully cloned. The positive ES clone gene in pPIC9 expression vector was sieved, and its coding sequence was identified to be as same as the previously reported sequence.
Conclusion 　The successful construction of ES gene in pPIC9 expression vector using molecular biological method maybe helpful for the high expression of ES protein, which may lay the foundation for the treatment of malignant tumor through anti-angiogenesis appoach.

Citation： ZHOU Dongfeng,ZHANG Zhongguang. Construction of Human Endostatin in Yeast Eukaryotic Expression Vector. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2007, 14(4): 417-419. doi: Copy

1.	O’Reilly MS, Boehm T, Shing Y, et al. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth [Ｊ]　. Cell, 1997; 88(2)∶277.
2.	Davis DW, Shen Y, Mullani NA, et al. Quantitative analysis of biomarkers defines an optimal biological dose for recombinant human endostatin in primary human tumors [Ｊ]　. Clin Cancer Res, 2004; 10(1 Pt 1)∶33.
3.	Capillo M, Mancuso P, Gobbi A, et al. Continuous infusion of endostatin inhibits differentiation, mobilization, and clonogenic potential of endothelial cell progenitors [Ｊ]　. Clin Cancer Res, 2003; 9(1)∶377.
4.	贺祥，潘卫，黄耀诚，等. 血管生成抑制素Endostatin研究新进展 [Ｊ]　. 中国肿瘤生物治疗杂志， 2000； 7（1）∶76.
5.	杨林，王金万，汤仲明，等. 重组人血管内皮抑制素Ⅰ期临床研究 [Ｊ]　. 中国新药研究， 2004； 13（6）∶538.
6.	Boehm T, Folkman J, Browder T, et al. Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance [Ｊ]　. Nature, 1997; 390(6658)∶404.
7.	Nyberg P, Heikkila P, Sorsa T, et al. Endostatin inhibits human tongue carcinoma cell invasion and intravasation and blocks the activation of matrix metalloprotease-2, -9, and -13 [Ｊ]　. J Biol Chem, 2003; 278(25)∶22404.
8.	刘然义，易继林，邵俊伟，等. 内皮抑素转基因治疗裸鼠人肝癌的实验研究[Ｊ]. 中国普外基础与临床杂志， 2006； 13（2）∶145.
9.	Hansma AH, Broxterman HJ, van der Horst I, et al. Recombinant human endostatin administered as a 28-day continuous intravenous infusion, followed by daily subcutaneous injections: a phase Ⅰ and pharmacokinetic study in patients with advanced cancer [Ｊ]　. Ann Oncol， 2005; 16(10)∶1695.
10.	Minning S, Serrano A, Ferrer P, et al. Optimization of the high-level production of Rhizopus oryzae lipase in Pichia pastoris [Ｊ]　. J Biotechnol， 2001; 86(1)∶59.
11.	Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris [Ｊ]　. FEMS Microbiol Rev， 2000; 24(1)∶45.

1. 　O’Reilly MS, Boehm T, Shing Y, et al. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth [Ｊ]　. Cell, 1997; 88(2)∶277.
2. 　Davis DW, Shen Y, Mullani NA, et al. Quantitative analysis of biomarkers defines an optimal biological dose for recombinant human endostatin in primary human tumors [Ｊ]　. Clin Cancer Res, 2004; 10(1 Pt 1)∶33.
3. 　Capillo M, Mancuso P, Gobbi A, et al. Continuous infusion of endostatin inhibits differentiation, mobilization, and clonogenic potential of endothelial cell progenitors [Ｊ]　. Clin Cancer Res, 2003; 9(1)∶377.
4. 　贺祥，潘卫，黄耀诚，等. 血管生成抑制素Endostatin研究新进展 [Ｊ]　. 中国肿瘤生物治疗杂志， 2000； 7（1）∶76.
5. 　杨林，王金万，汤仲明，等. 重组人血管内皮抑制素Ⅰ期临床研究 [Ｊ]　. 中国新药研究， 2004； 13（6）∶538.
6. 　Boehm T, Folkman J, Browder T, et al. Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance [Ｊ]　. Nature, 1997; 390(6658)∶404.
7. 　Nyberg P, Heikkila P, Sorsa T, et al. Endostatin inhibits human tongue carcinoma cell invasion and intravasation and blocks the activation of matrix metalloprotease-2, -9, and -13 [Ｊ]　. J Biol Chem, 2003; 278(25)∶22404.
8. 　刘然义，易继林，邵俊伟，等. 内皮抑素转基因治疗裸鼠人肝癌的实验研究[Ｊ]. 中国普外基础与临床杂志， 2006； 13（2）∶145.
9. 　Hansma AH, Broxterman HJ, van der Horst I, et al. Recombinant human endostatin administered as a 28-day continuous intravenous infusion, followed by daily subcutaneous injections: a phase Ⅰ and pharmacokinetic study in patients with advanced cancer [Ｊ]　. Ann Oncol， 2005; 16(10)∶1695.
10. 　Minning S, Serrano A, Ferrer P, et al. Optimization of the high-level production of Rhizopus oryzae lipase in Pichia pastoris [Ｊ]　. J Biotechnol， 2001; 86(1)∶59.
11. 　Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris [Ｊ]　. FEMS Microbiol Rev， 2000; 24(1)∶45.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Construction of Human Endostatin in Yeast Eukaryotic Expression Vector

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