Real-time quantitative PCR for evaluating murine thymic function / 南方医科大学学报
Journal of Southern Medical University
; (12): 62-65, 2006.
Article
in Zh
| WPRIM
| ID: wpr-234195
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To establish a real-time quantitative PCR method for detecting the levels of the signal joint T cell receptor excision circles (sjTRECs) in murine thymocytes and spleen lymphocytes for determining the amount of naive T cells and evaluating the thymic function.</p><p><b>METHODS</b>The genomic DNA was extracted from murine thymocytes and splenocytes for PCR amplification of the target fragments. After purification of the PCR product, the recombination-activating gene 2 (RAG(2)) fragment was cloned into pGEMT-Easy vector to construct the standard plasmid. After PCR optimization, the standard curve was obtained and the samples (thymocytes and splenocytes of BALB/c and C(57)BL/6 mice) were detected for sjTRECs by real-time quantitative PCR.</p><p><b>RESULTS</b>The standard plasmid was correctly constructed, and the standard curve with high reliability was obtained. No statistical difference was observed in sjTREC contents in the T lymphocytes between the two mouse strains.</p><p><b>CONCLUSIONS</b>Real-time quantitative PCR for sjTREC analysis is established successfully, which offers an important means for thymic function analysis and a reliable model establishment for study the thymus.</p>
Full text:
1
Index:
WPRIM
Main subject:
Thymus Gland
/
Receptors, Antigen, T-Cell
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T-Lymphocytes
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Gene Rearrangement, T-Lymphocyte
/
Polymerase Chain Reaction
/
Lymphocyte Count
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Cell Biology
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Allergy and Immunology
/
Genetics
/
Metabolism
Limits:
Animals
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2006
Type:
Article