Bacteroides fragilis and applications thereof
A technology of Bacteroides fragilis and strains, which is applied in the fields of food and daily chemical products, medicines, microorganisms, and health care products, can solve the problems of weak clinical curative effect, and achieve the effect of remarkable curative effect and strong stomach acid ability
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Embodiment 1
[0041] Isolation and purification of Bacteroides fragilis ZY-312
[0042] Reagents and instruments:
[0043] (1) Medium A: Bacteroides-bile-escin (BBE) agar (Qingdao Haibo Biotechnology Co., Ltd., product number: HB7028) was added with an improved formula, and the specific components are shown in Table 1:
[0044] Table I
[0045]
[0046] (2) Medium B: Bacteroides-bile-escin (BBE) agar (Qingdao Haibo Biotechnology Co., Ltd., product number: HB7028) was added with an improved formula, and the specific ingredients are as follows in Table 2:
[0047] Table II
[0048]
[0049] (3) Medium C: Fetal bovine serum was added to Brinell Broth (Qingdao Haibo Biotechnology Co., Ltd., product number: HB0241), and the addition amount was 5% (v / v) (Zhejiang Tianhang Biotechnology Co., Ltd., Brand: Sijiqing, Item No.: HB0205).
[0050] (4) Experimental equipment
[0051] 2.5L sealed culture tank (Mitsubishi Gas Chemical Co., Ltd., C-31)
[0052] Constant temperature incubator (Sh...
Embodiment 2
[0090] Identification of Bacteroides fragilis ZY-312
[0091] The polymerase chain reaction (PCR) primer (synthesized by Yingwei Jieji (Shanghai) Trading Co., Ltd.) sequence is as follows:
[0092] Primer pair 1:
[0093] Forward primer: 5'-ACGCTTGCACCCCTCCGTATTA-3'
[0094] Reverse primer: 5'-GCTTAGAGTTTGATCCTGGCTCAG-3'
[0095] Primer pair 2:
[0096] Forward primer: 5'-TGGGTGGTTGCTGCCTGGACACA-3'
[0097] Reverse primer: 5'-CATCCGGGTATGGATATGAA-3'
[0098] Primer pair 3:
[0099] Forward primer: 5'-GATGCTCCAGTTACAGCTTCCATTG-3'
[0100] Reverse primer: 5'-CGCCCAGTATATGACCTAGTTCGTG-3'
[0101] bft gene primer pair:
[0102] Forward primer: 5'-GACGGTGTATGTGATTTGTCTGAGAGA-3'
[0103] Reverse primer: 5'-ATCCCTAAGATTTATTATCCCAAGTA-3'
[0104] 1. PCR identification (PCR is the polymerase chain reaction, which is a commonly used method for rapidly amplifying genes)
[0105] (1) 16S rRNA sequence determination
[0106] The above strains were inoculated on medium A, and cult...
Embodiment 3
[0128] Tolerance of Bacteroides fragilis ZY-312 to Gastric Acid
[0129] (1) Artificial gastric juice preparation (according to the 2010 "Chinese Pharmacopoeia" artificial gastric juice preparation method)
[0130] Dissolve 23.4mL of concentrated HCl in 100mL of purified water to obtain dilute hydrochloric acid. Take 8.2mL dilute hydrochloric acid, add 400mL purified water and 5g pepsin ( Porcine source, 1:15000), dilute to 500mL. Stir overnight at 37°C with magnetic force to obtain artificial gastric juice.
[0131] (2) Cell preparation
[0132] Collect the bacterial solution, centrifuge, discard the supernatant, resuspend in normal saline, centrifuge again, discard the supernatant, and keep the bacteria for later use.
[0133] (3) adding artificial gastric juice to determine the number of viable bacteria
[0134] Add artificial gastric juice to the bacteria, resuspend, and measure the number of viable bacteria at 0, 1, 2, and 3 hours respectively.
[0135] The 10-fold...
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