Preparation of polyclonal antibody of recombinant human thioredoxin-1 and its protective effects on neonatal rats with endotoxemia / 中国当代儿科杂志
Chinese Journal of Contemporary Pediatrics
; (12): 837-841, 2011.
Article
在 Zh
| WPRIM
| ID: wpr-272456
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To clone the gene human thioredoxin 1 (hTrx-1) expressing its protein in the E.coli expression system and to obtain its polyclonal antibody, and to study the protective effects of hTrx-1 on neonatal rats with endotoxemia.</p><p><b>METHODS</b>DNA encoding hTrx-1 from fetal liver cells was isolated by RT-PCR. The hTrx-1 was cloned to the prokaryotic expression plasmid PET-28a to induce its protein expression in the E.coli expression system. The purified hTrx-1 was injected into rats to prepare polyclonal antibody. Newborn Sprague-Dawley rats were randomly assigned to three groups: control, lipopolysaccharide (LPS) and hTrx-1 (n=12 each). The control and the LPS groups were intraperitoneally injected with normal saline and LPS (5 mg/kg), respectively. The hTrx-1 group received an intraperitoneal injection of hTrx-1 (10 mg/kg) 30 minutes before LPS injection. The mortality rate 24 hrs after injection was compared between the three groups.</p><p><b>RESULTS</b>The prokaryotic expression plasmid PET-28a-hTrx-1 was constructed. The hTrx-1 protein was expressed and purified. The polyclonal antibody of hTrx-1 with the titer of 1∶51200 was prepared. The mortality rate of the control, LPS and hTrx-1 groups was 0, 67% and 17%, respectively (χ2=14.400, P<0.01).</p><p><b>CONCLUSIONS</b>The polyclonal antibody of hTrx-1 is prepared successfully. The hTrx-1 protein has protective effects on neonatal rats with endotoxiamia.</p>
全文:
1
索引:
WPRIM
主要主题:
Thioredoxins
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Recombinant Proteins
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Enzyme-Linked Immunosorbent Assay
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Blotting, Western
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Rats, Sprague-Dawley
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Endotoxemia
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Therapeutic Uses
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Allergy and Immunology
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Genetics
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Animals, Newborn
限制:
Animals
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Humans
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Male
语言:
Zh
期刊:
Chinese Journal of Contemporary Pediatrics
年:
2011
类型:
Article