Inhibitor of Neutrophil Proteinases　Elafin Expression in Pichia pastoris and Activity Investigation

China Biotechnology

研究报告

Inhibitor of Neutrophil Proteinases　Elafin Expression in Pichia pastoris and Activity Investigation

Download:

HTML

PDF(0KB) HTML
Export: BibTeX | EndNote (RIS)

Abstract Elafin is a specific inhibitor of neutrophil elastase.To establish expression system of neutrophil proteinases inhibitor elafin in Pichia pastoris ， Elafin cDNA was obtained by reverse transcription PCR(RT-PCR).With restricted enzyme digestion,gene of interest was cloned into shuttle vector pPIC9K.Then the construct was linearized and transformed into Pichia pastoris competent cells by electroporation.Selected positive clone was inoculated into buffered minimal glycerol(BMG)culture media.Add 100% methanol to a final concentration of 1.0% every 24 hours to maintain induction.At several different time points, transfer 1ml of expression cultrure to centrifuge to analyze the expression level by Coomassie-stained SDS-PAGE.Concentrated supernatnat was loaded onto a S6 column using a chromatographic system to purify secreted reconbinant elafin(rec-elafin).Rec-elafin immunogen and activity against proteinases was investigated by western blot and elastin-PAGE respectively. The rec-elafin expression level was up to 12 mg/L.Secreted elafin in Pichia pastoris was easily purified by one-step cationic exchange chromatography and retained its activity.

Key words： expression purification Pichia pastori neutrophil proteinases inhibitor

Received: 26 June 2006 Published: 25 December 2006

	Service
	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors

Cite this article:

. Inhibitor of Neutrophil Proteinases　Elafin Expression in Pichia pastoris and Activity Investigation. China Biotechnology, 2006, 26(12): 29-33.

URL:

https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I12/29

[1]	QIAO Sheng-tai,WANG Man-qi,XU Hui-ni. Functional Analysis of Prokaryotic Expression Protein of Tomato SlTpx in Vitro[J]. China Biotechnology, 2021, 41(8): 25-32.

[2]	ZHANG Ling,CAO Xiao-dan,YANG Hai-xu,LI Wen-lei. The Application of Continuous Purification in Affinity Chromatography and Evaluation of Production Scale-up[J]. China Biotechnology, 2021, 41(6): 38-44.

[3]	LI Bing,ZHANG Chuan-bo,SONG Kai,LU Wen-yu. Research Progress in Biosynthesis of Rare Ginsenosides[J]. China Biotechnology, 2021, 41(6): 71-88.

[4]	WANG Hui-lin,ZHOU Kai-qiang,ZHU Hong-yu,WANG Li-jing,YANG Zhong-fan,XU Ming-bo,CAO Rong-yue. Research Progress of Human Coagulation Factor VII and the Recombinant Expression Systems[J]. China Biotechnology, 2021, 41(2/3): 129-137.

[5]	ZHANG Lei,TANG Yong-kai,LI Hong-xia,LI Jian-lin,XU Yu-xin,LI Ying-bin,YU Ju-hua. Advances in Promoting Solubility of Prokaryotic Expressed Proteins[J]. China Biotechnology, 2021, 41(2/3): 138-149.

[6]	CHEN Zhong-wei,ZHENG Pu,CHEN Peng-cheng,WU Dan. Screening and Characterization of Thermostable Phytase Mutants[J]. China Biotechnology, 2021, 41(2/3): 30-37.

[7]	LIU Mei-qin,GAO Bo,JIAO Yue-ying,LI Wei,YU Jie-mei,PENG Xiang-lei,ZHENG Yan-peng,FU Yuan-hui,HE Jin-sheng. Long Non-coding RNA Expression Profile in A549 Cells Infected with Human Respiratory Syncytial Virus[J]. China Biotechnology, 2021, 41(2/3): 7-13.

[8]	YANG Xi,LUAN Yu-shi. Preliminary Study of Sly-miR399 in Tomato Resistance to Late Blight[J]. China Biotechnology, 2021, 41(11): 23-31.

[9]	CHEN Su-fang,XIA Ming-yin,ZENG Li-yan,AN Xiao-qin,TIAN Min-fang,PENG Jian. Recombinant Expression and Detection of Antimicrobial Activity of Cec4a[J]. China Biotechnology, 2021, 41(10): 12-18.

[10]	CHEN Xin-jie,QIAN Zhi-lan,LIU Qi,ZHAO Qing,ZHANG Yuan-xing,CAI Meng-hao. *Modification of Aromatic Amino Acid Synthetic Pathway in Pichia pastoris* to Produce Cinnamic Acid and ρ-Coumaric Acid**[J]. China Biotechnology, 2021, 41(10): 52-61.

[11]	SHI Peng-cheng, JI Xiao-jun. Advances in Expression of Human Epidermal Growth Factor in Yeast[J]. China Biotechnology, 2021, 41(1): 72-79.

[12]	RAO Hai-mi,LIANG Dong-mei,LI Wei-guo,QIAO Jian-jun,CAI YIN Qing-ge-le. Advances in Synthetic Biology of Fungal Aromatic Polyketides[J]. China Biotechnology, 2020, 40(9): 52-61.

[13]	DENG Tong,ZHOU Hai-sheng,WU Jian-ping,YANG Li-rong. Enhance Soluble Heteroexpression of a NADPH-Dependent Alcohol Dehydrogenase Based on the Chaperone Strategy[J]. China Biotechnology, 2020, 40(8): 24-32.

[14]	ZHANG Xiao-hang,LI Yuan-yuan,JIA Min-xuan,GU Qi. Identification and Expression of Elastin-like Polypeptides[J]. China Biotechnology, 2020, 40(8): 33-40.

[15]	LV Yi-fan,LI Geng-dong,XUE Nan,LV Guo-liang,SHI Shao-hui,WANG Chun-sheng. Prokaryotic Expression, Purification of LbCpf1 Protein Gene and in Vitro Cleavage Activity Assay[J]. China Biotechnology, 2020, 40(8): 41-48.

Viewed

Full text

Abstract

Cited

Shared

Discussed