Schizothorax prenanti HSP60 (heat shock protein 60) gene cDNA (complementary deoxyribonucleic acid) full-length sequence and application thereof
A technology of Schizothorax and its application, applied in the direction of genetic engineering, plant gene improvement, application, etc., can solve the problems of Schizothorax HSP60 and Schizothorax not seen, and reach the prospect of clinical application Good, damage-reducing effect
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[0029] Example 1 cDNA cloning of Hsp60 gene of Schizothorax
[0030] 1. Test materials
[0031] 1. Bacteria / carrier
[0032] Escherichiacoli (E.coli) DH5α competent cells (purchased from Tiangen Biochemical Technology Co., Ltd., Beijing, China); pMD19-T (purchased from Bao Bioengineering Co., Dalian, China).
[0033] 2. Experimental animals
[0034] Healthy Schizophrenia was purchased from Sichuan Runzhao Fishery Co., Ltd.
[0035] 3. Main reagents
[0036] (1) Sanprep column plasmid DNA small extraction kit, Sanprep column DNA gel recovery kit, sample RNA / DNA protectant, RNAiso Plus lysate, PrimeScript TM Reagent Kit (Perfect Real Time) cat#RR037A (purchased from Bao Biological Engineering Company, Dalian, China).
[0037] (2) 2×Taq Master Mix, Goldview, Agarose, DNA Marker (DL2000, DL1000DNAMarker) (purchased from Tiangen Biochemical Technology Co., Ltd., Beijing, China).
[0038] (3) SMART TM RACE cDNA Amplification Kit (purchased from BD Biosciences Clontech, Palo Alto, CA, USA)
[0...
Example Embodiment
[0144] Experimental example 1 The expression level of Hsp60 gene after Streptococcus agalactiae infection
[0145] 1. Test materials
[0146] 1. Bacteria
[0147] Streptococcus agalactiae (S. agalactiae) was provided by the Fish Disease Research Center of Sichuan Agricultural University.
[0148] 2. Main reagents
[0149] Prime Script TM Reagent Kit With gDNA Eraser(Perfect Real Time)cat#RR047A、 Premix Ex Taq TM II (Tli RNaseH Plus) (purchased from Bao Biological Engineering Company, Dalian, China); Blood total protein, total RNA, DNA and microRNA-kit (purchased from Beijing Bolingkewei Biotechnology Co., Ltd.).
[0150] 3. Main instruments
[0151] Step One Plus TM Real-time fluorescent quantitative PCR system-Life Tech (applied biosystems) (life-technologies company). Other instruments The instrument of Example 1.
[0152] 4. Experimental animals
[0153] Healthy Schizophrenia was purchased from Sichuan Runzhao Fishery Co., Ltd.
[0154] 5. Main reagent configuration
[0155] (1) Med...
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