Cloning, expression, purification and activity of the hsBLyS

Ping Liu, Shuang Quan Zhang, Xiao Mei Yan, Haojie Huang, Kai Ya Zhou

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The full length cDNA of human B lymphocyte stimulator (hBLyS) was amplified by using PCR method from cDNA library of human placenta. After purifying and sequencing, the DNA fragment of functional domain of hBLyS (hsDNA fragment) was amplified by using nested PCR method from the PCR product. The prokaryotic expression plasmid pET-30a(+)/hsBLyS was constructed with recombinant DNA techniques after purifying and identifying the hsDNA fragment. Then the plasmid pET-30a(+)/hsBLyS was transformed into λDE3 cells and the recombination protein was found to be highly expressed; the expression product was purified by affinity chromatography gel, Ni 2+-IDA, made in our laboratory. The experimental results showed that the sequence of the PCR product was identical with the published hBLyS cDNA sequence and purity of the recombination protein we obtained was high. The activity of the purified recombination protein was very significant in the proliferation test of B lymphocytes.

Original languageEnglish (US)
Pages (from-to)213-214
Number of pages2
JournalActa Biochimica et Biophysica Sinica
Volume33
Issue number2
StatePublished - 2001
Externally publishedYes

Fingerprint

B-Cell Activating Factor
Cloning
Purification
Organism Cloning
Genetic Recombination
Polymerase Chain Reaction
Plasmids
Complementary DNA
Affinity chromatography
Proteins
Recombinant DNA
Lymphocytes
Gene Library
Gels
DNA Sequence Analysis
Affinity Chromatography
Placenta
B-Lymphocytes
DNA

Keywords

  • Cell apoptosis
  • Cell proliferation
  • Fusion protein
  • HsBLyS
  • Transmembrane protein

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

Liu, P., Zhang, S. Q., Yan, X. M., Huang, H., & Zhou, K. Y. (2001). Cloning, expression, purification and activity of the hsBLyS. Acta Biochimica et Biophysica Sinica, 33(2), 213-214.

Cloning, expression, purification and activity of the hsBLyS. / Liu, Ping; Zhang, Shuang Quan; Yan, Xiao Mei; Huang, Haojie; Zhou, Kai Ya.

In: Acta Biochimica et Biophysica Sinica, Vol. 33, No. 2, 2001, p. 213-214.

Research output: Contribution to journalArticle

Liu, P, Zhang, SQ, Yan, XM, Huang, H & Zhou, KY 2001, 'Cloning, expression, purification and activity of the hsBLyS', Acta Biochimica et Biophysica Sinica, vol. 33, no. 2, pp. 213-214.
Liu, Ping ; Zhang, Shuang Quan ; Yan, Xiao Mei ; Huang, Haojie ; Zhou, Kai Ya. / Cloning, expression, purification and activity of the hsBLyS. In: Acta Biochimica et Biophysica Sinica. 2001 ; Vol. 33, No. 2. pp. 213-214.
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