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AssigneeChui Chi LAM
InventorsWan Keung Raymond Wong, Xiu Hua Hu, Ka Lun Ng, Chui Chi Lam
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The present invention is concerned with a method of production of authentic and bioactive human basic fibroblast growth factor (bFGF) of 146 amino acids and without any modification at either C- or N-terminal of the bFGF. The method includes the steps as of providing a Bacillus subtilis host, introducing a DNA construct into the Bacillus subtilis host to produce a transformed Bacillus subtilis host, the DNA construct including an insert consisting of, from 5′ to 3′, a cellulose binding domain (CellBD), an intein sequence and a DNA coding for the bFGF polypeptide, and subjecting the transformed Bacillus subtilis host to a shake flask cultivation process or a fed-batch fermentation process.
A method of production of authentic and bioactive human basic fibroblast growth factor (bFGF) of 146 amino acids intracellularly and without any modification at either C- or N-terminal of the bFGF, comprising the steps of providing a Bacillus subtilis host and introducing a DNA construct into the Bacillus subtilis host to produce a transformed Bacillus subtilis host, wherein the DNA construct including an insert having, from 5′ to 3′, a cellulose binding domain (CellBD), an intein sequence and a DNA coding for the bFGF polypeptide, and subjecting the transformed Bacillus subtilis host to a shake flask cultivation process or a fed-batch fermentation process, whereby to enable the transformed Bacillus subtilis host to produce the bFGF in a soluble form, cleaved, independent from proteins encoded by DNA regions preceding and subsequent to the bFGF DNA coding in the insert, and intracellularly.
A biological system engineered from a Bacillus subtilis host, comprising a DNA construct including an insert comprising, from 5′ to 3′, a cellulose binding domain (CellBD), an intein sequence and a DNA coding for the bFGF polypeptide.
A DNA construct comprising an insert comprising, from 5′ to 3′, a cellulose binding domain (CellBD), an intein sequence and a DNA coding for a bFGF polypeptide, wherein the bFGF polypeptide is an authentic and bioactive human basic fibroblast growth factor of 146 amino acids intracellularly and without any modification at either C- or N-terminal of the bFGF.
Dependencies are inferred from the claim text.