Severity: Warning
Message: file_get_contents(https://...@pubfacts.com&api_key=b8daa3ad693db53b1410957c26c9a51b4908&a=1): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests
Filename: helpers/my_audit_helper.php
Line Number: 176
Backtrace:
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 176
Function: file_get_contents
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 250
Function: simplexml_load_file_from_url
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 1034
Function: getPubMedXML
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 3152
Function: GetPubMedArticleOutput_2016
File: /var/www/html/application/controllers/Detail.php
Line: 575
Function: pubMedSearch_Global
File: /var/www/html/application/controllers/Detail.php
Line: 489
Function: pubMedGetRelatedKeyword
File: /var/www/html/index.php
Line: 316
Function: require_once
Aim: To clone, express, and identify a fragment of Cor h 1 from Corylus heterophylla.
Methods: Through bioinformatics predication, the antigenic epitope of Cor h 1 was selected. A fragment gene of Cor h 1 was amplified by PCR and cloned into pMD18-T vector for sequencing. Then the fragment gene was sub cloned into pET-32a expression vector for expression, and then purified by metal (Ni(2+);) chelating affinity chromatography. The immunogenicity was tested by Western blot.
Results: The length of the fragment gene was 243 bp, coding 81 amino acids; the relative molecular mass of recombinant protein was 9 000. And the fragment of Cor h 1 was mainly expressed as soluble protein, purified protein has good immunogenicity.
Conclusion: The fragment gene of Cor h 1 was successfully cloned and expressed in this study, and the recombinant protein possessed good IgE-binding capacity.
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