Biomedical subjects

Zhe-Yu Chen

Publications and source records attributed to Zhe-Yu Chen.

24 records · Page 2Linked to original sources

PubMed2000

A Structure-function Analysis of Human GDNF.

The glial cell line-derived neurotrophic factor(GDNF) plays a very important role in the regeneration of the nervous system. Based on the results of the X-ray structure analysis of rat GDNF, human GDNF gene was modified with deletion and insertion mutagenesis by using PCR methods. The various mutants were all highly expressed in E.coli. The recombinant proteins were purified and their survival-promoting activities were determined by using cultures of the spinal cord neurons of embryonic mouse. The results showed that the "cystine knot motif" was critical for the maintenance of GDNF structure the alpha-helix, finger 1 and finger 2 region were critical for GDNF neurotrophic activity and the N-terminus of human GDNF was not essential for its biological functions.

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PubMed2000

High Expression of Human Persephin in Insect Cells.

The human persephin (PSP) was expressed in Tn-5B1-4 insect cells using the Bac to Bac baculovirus expression system. The expressed product amounted for 20% of total cellular soluble proteins. The expressed product was purified by Ni(2+) affinity chromatography and the activity assays showed that it could significantly prolong the survival of spinal cord neurons.

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PubMed1999

Cloning and Expression of Human Manganese-superoxide Dismutase cDNA.

The cDNA encoding human manganese-superoxide dismutase was amplified from the human liver cell (L02) total RNA by RT-PCR and sequenced. The human Mn-SOD cDNA was ligated into expression vector pET-24a(+) under T7 promoter. After 5 h induction with 1 mmol/L IPTG 800 mol/L Mn(2+) human Mn-SOD was highly expressed in E.coli BL21(DE3). The protein product was up to 50% of the bacteria total protein in soluble form and had specific SOD activity.

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PubMed1999

Cloning of Human Glial Neurotrophic Factor and Its High Expression in Insect Cells.

The human glial cell derived neurotrophic factor (GDNF) was expressed in Tn-5B1-4 insect cells using the Bac to Bac baculovirus expression system, accounting for 30% of total cellular soluble proteins. The expressed product was purified by affinity chromatography and the activity assays showed that it could significantly promote the survival of dopaminergic neurons. The study can be the basis for further structural-functional analysis of GDNF.

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PubMed1999

Construction of Biotinylated Annexin V for Detection of Apoptosis.

The biotinylated annexin V was generated by genetic engineering method. The encoding region of annexin V was fused with a gene coding for the carboxyl terminal 87 residues of Escherichia coli biotin carboxyl carrier protein. The fused gene was expressed in E.coli and the annexin V was biotinylated in vivo. The biotinylation efficiency was about 60% as detected by HPLC. The biotinylated annexin V was purified by avidin affinity chromatography and used to detect the apoptosis of the neurons induced by morphine.

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PubMed1999

The Human Recombinant GDNF and Study of Its Biological Activity.

The cDNA encoding the mature human glial cell derived neurotrophic factor(GDNF) was isolated by using RT-PCR method from total RNA extracted of fetal human brain. The expression plasmid pET-GDNF was constructed by inserting GDNF cDNA into plasmid pET-28a(+) containing T7 promoter and transformed into E.coli BL21(DE3). An expression strain BLGDNF was selected. SDS-PAGE analysis revealed that the human GDNF protein was highly expressed and accumulated up to above 30% of the total bacterial proteins in the form of inclusion body after the induction. The antibody to GDNF was prepared by immunization of rat using purified GDNF protein. Purified and refolded GDNF protein could significantly promote the survival of dopaminergic neurons.

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