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Application of signal recognition particle subunit and encoding gene thereof to preparation of acid-resistant bifidobacterium, plasmid containing gene and bifidobacterium

A technology of signal recognition and bifidobacteria, applied in the field of genetic engineering, can solve problems such as poor taste, unstable genetic characteristics of bifidobacteria, and cumbersome operations

Active Publication Date: 2016-06-15
BEIJING UNIV OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the coating process is cumbersome, and the taste of yogurt or lactic acid beverages added with coated strains will deteriorate, and the genetic characteristics of acid-resistant and domesticated bifidobacteria are also relatively unstable.

Method used

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  • Application of signal recognition particle subunit and encoding gene thereof to preparation of acid-resistant bifidobacterium, plasmid containing gene and bifidobacterium
  • Application of signal recognition particle subunit and encoding gene thereof to preparation of acid-resistant bifidobacterium, plasmid containing gene and bifidobacterium
  • Application of signal recognition particle subunit and encoding gene thereof to preparation of acid-resistant bifidobacterium, plasmid containing gene and bifidobacterium

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Experimental program
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Effect test

Embodiment 1

[0048] 1. Recombinant expression plasmid P 18 Construction of P-Ffh

[0049] (1) Construction of expression plasmid P 18 P

[0050] The pDG-7 plasmid and the pUC18 plasmid were double-digested with BamHI and HindIII respectively, and the pMB1 of the pDG-7 digestion product was recovered, and the pUC18 digestion product was recovered, and ligated in the T4 DNA ligase reaction system at 4°C for 10-12 hours. Transform the plasmid into DH5α Escherichia coli competent cells by heat shock, plate on LB solid medium containing 100 μg / mL ampicillin and culture for 16 hours, pick a single colony in LB liquid medium containing 100 μg / mL ampicillin After enrichment and extraction, a 4758bp plasmid P was obtained 18 P( Figure 1B ).

[0051] (2) Preparation of gene encoding signal recognition particle subunit

[0052] Use the upstream primer (PrimerF:CG as shown in SEQIDNo: 4 GAATTC CTTGACAGACAGACTCTCGAATGCGT, the underline is the EcoRI restriction site) and the downstream primer s...

Embodiment 2

[0079] Each step was carried out according to the method of Example 1, except that the bifidobacterium used was Bifidobacterium juvenile CGMCC No.6270.

[0080] The results show that the conversion has P 18 The survival rates of this strain of P-Ffh at pH 1, 2 and 3.5 were 2%, 5% and 8%, respectively, and the survival rates of normal strains at pH 1, 2 and 3.5 were 0.1%, 0.7% and 1.5%, respectively.

Embodiment 3

[0082] Carry out each step according to the method of embodiment 1, difference is, the plasmid used is not P 18 P, but the pMG36e plasmid.

[0083] The results showed that the expression level of the plasmid in CGMCCNo.2265 was extremely low, and could only be vaguely seen in agarose electrophoresis, and the survival rates of CGMCCNo.2265 transformed with the plasmid were 3%, 3%, and 3.5% respectively in pH1, 2, and 3.5. 5% and 7%.

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Abstract

The invention relates to the field of bioengineering, particularly discloses a signal recognition particle subunit shown in SEQ ID No:1 and / or application of a nucleotide sequence encoding the signal recognition particle subunit to preparation of acid-resistant bifidobacterium, and further discloses a recombinant expression plasmid containing the nucleotide sequence and the acid-resistant bifidobacterium containing the signal recognition particle subunit and / or the nucleotide sequence encoding the signal recognition particle subunit and / or the acid-resistant bifidobacterium of the recombinant expression plasmid. According to the technical scheme, a gene encoding the signal recognition particle subunit shown in SEQ ID No:1, preferentially a plasmid P18P containing the gene is converted into bifidobacterium which is preferred as bifidobacterium CGMCCNo.2265, and the gene can resistant to acid with the pH being 1, the preferential pH being 2 and the optimal pH being 3.5.

Description

technical field [0001] The present invention relates to the field of genetic engineering, in particular to the application of the signal recognition particle subunit shown in SEQIDNo: 1 and / or the nucleotide sequence encoding the signal recognition particle subunit in the preparation of acid-resistant bifidobacteria, and the Nucleotide sequences for recombinant expression plasmids and acid fast bifidobacteria. Background technique [0002] Bifidobacteria are the most beneficial flora in the intestine, and the suitable pH value for growth is 6.5-7.5. The reduction or even disappearance of the number of bifidobacteria is a sign of an "unhealthy" state. Bifidobacteria are a barometer of human health. The main way to replenish bifidobacteria is to drink yogurt or beverages containing bifidobacteria, or directly drink the probiotics of bifidobacteria (bacteria powder or 1:3 serving liquid). [0003] It is known that the acidity of human gastric juice is generally 0.9-1.5. Under ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/31C12N15/74C12R1/01
CPCC07K14/195C12N15/74C12N2800/101
Inventor 金君华任发政王芳张红星刘慧马京升龚煜欣李雪
Owner BEIJING UNIV OF AGRI
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