کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2021111 | 1069228 | 2009 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: The production of recombinant human laminin-332 in a Leishmania tarentolae expression system The production of recombinant human laminin-332 in a Leishmania tarentolae expression system](/preview/png/2021111.png)
Laminin (LM)-332 (α3β3γ2), a large heterotrimeric glycoprotein, is an essential component of epithelial basement membranes that promotes cell adhesion and migration. Here, we expressed human LM-332 using a novel protein expression system based on the trypanosomatid protozoan host Leishmania tarentolae. Plasmids containing cDNA encoding full-length β3 and γ2 subunits and truncated α3 subunit were sequentially introduced into L. tarentolae. A recombinant strain harboring the three subunits of human LM-332 efficiently formed heterotrimer and secreted it into the culture medium. Heterotrimeric recombinant LM-332 (rLM-332) could be purified from culture medium with one-step immuno-affinity chromatography. The eluted fraction contained all three subunits, as confirmed by immunoprecipitation and immunoblotting. The purified rLM-332 showed similar cell adhesion activity to rLM-332 purified from mammalian cells, indicating its proper folding and assembly. The obtained expression level was not high; however, we suggest that this expression system has the potential for mass production of LMs for tissue engineering.
Journal: Protein Expression and Purification - Volume 68, Issue 1, November 2009, Pages 79–84