Bacterial Expression System
Bacillus subtilis has been widely used for extracellular production of native and homologous proteins. Based on the use of a synthetic expression/secretion cassette, a new bacterial expression system is developed and may be applied to produce heterologous proteins efficiently as secreted products.
This bacterial expression system is particularly for the expression of heterologous proteins for use with gram-positive and gram-negative bacteria, particularly Bacillus subtilis and E. coli. It comprises of novel genetic constructs. Also provided is a novel promoter element. The novel expression cassette, Veg cassette, allows very high expression of heterologous proteins to be achieved in B. subtilis. The cassette may be cloned into a modified B. subtilis/E. coli shuttle vector pRB373M2 to form an expression/secretion vector named the veg vector. The application of the cassette has successfully promote efficient secretory production of two widely different heterologous proteins, an endoglucanase (Eng) of Cellulomonas fimi and human epidermal growth factor (hEGF) in B. subtilis. Moreover, the cassette was successfully used to express and secrete proteins that were found to be difficult to produce in B. subtilis, e.g. hEGF.
1. High stability
2. High levels of extracellular products provided
3. Efficient production of protocols and clones of the two recombinant proteins (C. fimi Eng and hEGF)
4. Avoid the use of protease-deficient strains which are usually slow growing and difficult to handle
5. Render the heterologous gene cassette to be protected from the restriction of the host bacteria
- Production of secretory heterologous proteins in Bacillus subtilis and E. coli
- Production of large quantity of hormones, growth factors and peptide drugs
Patent Number: US6146848A
Application Number: US1998121286A
Inventor: Wong, Wan Keung | Lam, Kat Hon
Priority Date: 23 Jul 1998
Priority Number: US6146848A
Application Date: 23 Jul 1998
Publication Date: 14 Nov 2000
IPC Current: C07K0014485 | C12N001570 | C12N001574
US Class: 4350691 | 4352523 | 43525231 | 43525233 | 4353201 | 435471 | 5360241
Assignee Applicant: The Hong Kong University of Science & Technology
Title: Bacterial expression system
Usefulness: Bacterial expression system
Summary: (I) is useful for transforming bacteria which are useful for producing expression products such as heterologous proteins.
Novelty: Novel endonuclease-protected VegI promoter useful for producing heterologous protein
化工/材料
化工/材料应用
23 Jul 1998
US 6146848
TTC.PA.081
香港
