Promoter and use thereof
AssigneeAzenta US, Inc.
InventorsGaoxu Xue, Tianming Qi, Aihua Feng, Zhengli Xie, Yankai Jia, Xin Wu, Zhongping Sun, Guojuan Liao
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Abstract
An improved promoter and a use thereof. An improvement is to mutate a nucleic acid sequence between −35 region and −10 region in a promoter region into recognition sites for an endonuclease. The improvement is used for overcoming the problem that a transcription or translation product of foreign genes under a strong promoter might be toxic to a host and cannot be cloned and avoiding the phenomena of false positives and false negatives during blue-white screening.
Claims20
A β-galactosidase promoter, consisting of a −35 region, a −10 region, and a mutated nucleic acid sequence between the −35 region and the −10 region, wherein:
- the mutated nucleic acid sequence comprises at least one recognition site for an endonuclease, each recognition site for an endonuclease selected from the group consisting of EcoRV, AleI, BamHI, XhoI, and PmlI;
- the −35 region has a nucleic acid sequence of 5′-TTTACA-3′;
- a 5′ end of the mutated nucleic acid sequence is linked to a 3′ end of the −35 region;
- a 3′ end of the mutated nucleic acid sequence is linked to a 5′ end of the −10 region;
- and
- the mutated nucleic acid sequence is 16 to 18 base pairs in length.
Dependencies are inferred from the claim text.