Special-purpose medium for separating insect endophytic bacteria and preparation method thereof
A technology of endogenous bacteria and culture medium, applied in the field of bioengineering, can solve the problems that it is difficult to separate rare bacteria or uncommon bacteria resources, the endogenous bacteria of insects are not well targeted, and the original intention of the experiment cannot be achieved. Rich diversity, strong feasibility, and the effect of improving the bacteria yield
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Embodiment 1
[0022] The medium formula (1000ml) of embodiment 1 insect endophytic bacteria isolation culture:
[0023] Tenebrio molitor dry powder 2g, yeast powder 1g (G0961, Sangon Bioengineering (Shanghai) Co., Ltd.), gellan gum 3g (EZ0001, Shanghai Eka Biotechnology Co., Ltd.), trehalose 0.5g (TB0966, Sangon Bioengineering (Shanghai) Co., Ltd.), carrot juice 100ml (prepared from 300g red carrots), NaCl 9g (analytical pure, Xilong Chemical), agar powder 16g (01-023, Beijing Aoboxing Biotechnology Co., Ltd.), distilled water to volume To 1 000ml, natural pH.
Embodiment 2
[0024] The preparation process of embodiment 2 medium:
[0025] A. Preparation of Tenebrio molitor dry powder: Weigh an appropriate amount of mature larvae of Tenebrio molitor, dry at 50°C for 12 hours, grind with a mortar until there are no obvious particles, and pass through a 200-mesh sieve to obtain Tenebrio molitor dry powder.
[0026] B. Prepare carrot dipping juice: weigh 300g red carrot, cut into 2cm 3 After crushing the cube with a small pulverizer, transfer it to a conical flask, add 200ml of distilled water, 180r / min, shake and mix at room temperature for 20min. Finally, filter with four layers of gauze, and filter the filtrate for a second time with a sand core filter to obtain the required carrot dipping juice, which is made now and placed in an oven at 60°C to keep warm.
[0027] C. First weigh 5g of gellan gum, slowly dissolve it with 800ml of distilled water, then add 2g of Tenebrio molitor dry powder, 1g of yeast powder, 0.5g of trehalose, and 9g of NaCl, and...
Embodiment 3
[0029] Use the prepared medium to make a plate, and use the dilution of the intestinal homogenate of tussah silkworm Antheraea pernyi to coat the plate and culture it. Each sample is repeated 3 times. The obtained typical strains were identified by the combination of characteristics, physiological and biochemical index determination and 16S rRNA sequence alignment. The results show that the strains grow vigorously on the invented culture medium, the number is large, the reproducibility is strong, and the identified species are abundant. The list of main species is shown in Table 1. The endophytic bacteria in tussah silkworm gut reported in the literature mainly belong to Bacillus, Staphylococcus and Enterobacter. Therefore, compared with the traditional formula, the species of endophytic bacteria isolated by the formula of the present invention has obvious advantages in diversity.
[0030] Table 1 The intestinal endophytic bacteria of tussah silkworm isolated and purified by ...
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