Polypeptides and nucleic acids encoding same
A coding and nucleic acid sequence technology, applied in the field of polypeptides and nucleic acids encoding the polypeptides, can solve problems such as the potential utility of no amino acid sequences
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Embodiment 8
[0029] (i) an increase in the number of total facultative anaerobic bacteria in the body, for example in piglets and / or chicks, preferably ileo-rectal contents and / or cecal contents, respectively, in a medium supplemented with 5% vol / vol sheep blood Cultured on Brucella agar, incubated at 37°C in an anaerobic box for 5 days and then measured;
[0030] (ii) an increase in the number of Escherichia coli (Escherichia coli) in vivo, for example in piglets and / or chicks, preferably the ileo-rectal contents and / or cecal contents are respectively treated with Coli-ID pigment producing medium under aerobic conditions measured after incubation at 37°C for 24 hours.
[0031] (iii) The total lactic acid bacteria in the body are substantially unchanged or increased, for example, in piglets and / or chicks, preferably ileo-rectal contents and / or cecal contents, respectively, on MRS agar in an anaerobic box at 37°C Measured after 48 hours of incubation.
[0032] (iv) The total Lactobacillus...
Embodiment 6
[0092] The experiments of Example 6 were PCR reactions, performed in this example with DNA isolated from various strains of Bacillus licheniformis. In a specific embodiment of this assay, the DNA used as template for the PCR reaction is chromosomal DNA that can be isolated by techniques known in the art. The test of Example 6 was positive when a PCR fragment of the correct size was obtained. In Example 6, the correct size is expressed as 0.4kb. In a specific embodiment, the correct size is 0.35kb to 0.44kb (=350bp-440bp). In alternative embodiments, the correct size is 330-430bp, 340-420bp, 350-410bp, 360-400bp, 370-390bp or 385-395bp. The size of the coding sequence (CDS) of SEQ ID NO: 1 is about 380 bp (ie 378 bp).
[0093] nucleic acid sequence
[0094] The present invention also relates to an isolated nucleic acid sequence comprising a nucleic acid sequence encoding a polypeptide (a) encoding a polypeptide of the present invention; (b) hybridizing under very low string...
Embodiment 1
[0221] Example 1: Shake flask fermentation of Bacillus licheniformis
[0222] Bacillus licheniformis ATCC 14580 is propagated overnight at 37° C. on TY agar medium (TY culture fluid solidified with 2% agar), and inoculated into shake flasks comprising 100 ml of TY culture fluid. This TY culture fluid has the following components: Tryptone: 20g / l, yeast extract: 5g / l, FeCl 2 4H 2 O: 7mg / l, MnCl 2 4H 2 O: 1mg / l, MgSO 4 ·7H 2 O: 15 mg / l, pH 7.3. The shake flasks were incubated at 37°C for 20 hours with a shaking speed of 225 rpm. Cells were removed by centrifugation.
[0223] SDS polyacrylamide gel electrophoresis of the supernatant showed a band with a relative molecular weight of 12 kDa consistent with the expected polypeptide. The polypeptide of the present invention is called "L12 protein" because the relative molecular weight of the polypeptide measured by SDS-PAGE is 12 kDa (however, the theoretical molecular weight can be calculated to 9591.56 Da).
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