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[Construction of mic2/CD99 gene vector and its transfection in Hodgkin lymphoma L428 cell line].

Zuoping Huang, Ying He, Xinhua Zhou, Xi-qing Han, Zi-Qun Wu, Yan Zhang, Zonghua Wen, Tong Zhao

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Abstract

OBJECTIVE: To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells. METHODS: The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry. RESULTS: A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells. CONCLUSION: A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.

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What this paper is about

OBJECTIVE: To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells. METHODS: The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry. RESULTS: A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells. CONCLUSION: A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.

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Available abstract

OBJECTIVE: To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells. METHODS: The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry. RESULTS: A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells. CONCLUSION: A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.

Key concepts: Transfection, Complementary DNA, Molecular biology, Jurkat cells, Cell culture, Lipofectamine, Expression vector, Gene

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[Construction of mic2/CD99 gene vector and its transfection in Hodgkin lymphoma L428 cell line]. — Research Paper | ScholarLens