RESUMO
Endochitinase is a natural extracellular protein in Trichoderma longibrachiatum T6, which can degrade the eggshell of Heterodera avenae significantly, however the related genes that coding this protein was rarely characterized. In the present study, the endochitinase 18-5 gene (T6-Echi18-5) of T. longibrachiatum T6 was cloned and sequenced. The expression level of T6-Echi18-5 gene in T. longibrachiatum T6 was induced and increased after the H. avenae cysts inoculation. The full-length cDNA sequence of T6-Echi18-5 was 1671 bp that contained an ORF of 1275 bp, corresponding to 424 amino acids with a 45.9 kDa molecular weight. A single band of 60.04 kDa was detected and identified using SDS-PAGE and Western blot analysis after transferring the T6-Echi18-5 gene to Escherichia coli BL21 Rosetta (DE3). The concentration of purified recombinant T6-Echi18-5 protein was 1.53 mg·ml-1, and the optimal temperature and pH were 50 °C and 5.0, respectively. The eggshell and content were dissolved and exuded from 4 to10 days after treatment with the purified recombinant T6-Echi18-5 protein. The relative inhibition rate of eggs hatching was 86.79 % at 12 days after treatment. Our study demonstrated the key role of T6-Echi18-5 gene in degrading the H. avenae eggshell and inhibiting the eggs hatching.
Assuntos
Quitinases , Hypocreales , Trichoderma , Quitinases/genética , Trichoderma/metabolismo , Antinematódeos , Hypocreales/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
BACKGROUND: Coronary artery fistula refers to an abnormal communication between a coronary artery and great vessel, a cardiac chamber or other structure. The left circumflex artery (LCX) pericardia fistula combined with huge pseudoaneurysm is extremely rare. CASE PRESENTATION: A 39-year-old young female was admitted into our hospital because of palpitation and shortness of breath. Coronary computed tomography angiography (CCTA) showed a huge pseudoaneurysm located in pericardium. Coronary angiography revealed the LCX pericardia fistula. Then surgical treatment was performed. She was in good condition without complications after surgery. CONCLUSIONS: Coronary artery fistula combined with pseudoaneurysm can be caused by congenital factors. Early surgical treatment can relieve the patient's symptoms and prevent the occurrence of adverse cardiovascular events.