pcDPG parathyroid hormone gene therapy of hypoparathyroidism: an experimental study
To construct recombination eukaryote expression plasmid for human parathyroid hormone (PTH) gene, assay PTH expression and biological activity after transfection in vitro and evaluate gene therapy effect on hypoparathyroidism (HPT). (1) PTH gene was amplified from human embryonic parathyroid gland t...
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| Veröffentlicht in: | Zhong hua yi xue za zhi 2006-01, Vol.86 (4), p.260-265 |
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| container_title | Zhong hua yi xue za zhi |
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| creator | Wen, Ji-min Zhong, Yu-hua Sun, Yun Huang, Zhong Zhang, Ling-wu Wu, Quan-li Zeng, Da-cai |
| description | To construct recombination eukaryote expression plasmid for human parathyroid hormone (PTH) gene, assay PTH expression and biological activity after transfection in vitro and evaluate gene therapy effect on hypoparathyroidism (HPT).
(1) PTH gene was amplified from human embryonic parathyroid gland tissue, and plasmid pcDNA3.1-PTH-GFP (pcDPG) was constructed by TOPO recombination technique. Digestion, PCR and sequencing were used to identify the positive vectors. (2) pcDPG was transformed into 293 cells by Lipofectamine 2000(TM), fluorescent inverted microscope was used to observe GFP expression, and PTH gene expression was assayed by RT-PCR technique. (3) PTH protein in supernatant was purified and evaluated biological activity. (4) HPT rabbit models were developed and plasmid pcDPG was injected in skeletal muscles, respectively. Serum calcium, phosphonium and PTH were assayed and pathological changes observed.
(1) The findings in digestion and PCR were accorded to anticipation and sequences in report were id |
| format | Article |
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(1) PTH gene was amplified from human embryonic parathyroid gland tissue, and plasmid pcDNA3.1-PTH-GFP (pcDPG) was constructed by TOPO recombination technique. Digestion, PCR and sequencing were used to identify the positive vectors. (2) pcDPG was transformed into 293 cells by Lipofectamine 2000(TM), fluorescent inverted microscope was used to observe GFP expression, and PTH gene expression was assayed by RT-PCR technique. (3) PTH protein in supernatant was purified and evaluated biological activity. (4) HPT rabbit models were developed and plasmid pcDPG was injected in skeletal muscles, respectively. Serum calcium, phosphonium and PTH were assayed and pathological changes observed.
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(1) PTH gene was amplified from human embryonic parathyroid gland tissue, and plasmid pcDNA3.1-PTH-GFP (pcDPG) was constructed by TOPO recombination technique. Digestion, PCR and sequencing were used to identify the positive vectors. (2) pcDPG was transformed into 293 cells by Lipofectamine 2000(TM), fluorescent inverted microscope was used to observe GFP expression, and PTH gene expression was assayed by RT-PCR technique. (3) PTH protein in supernatant was purified and evaluated biological activity. (4) HPT rabbit models were developed and plasmid pcDPG was injected in skeletal muscles, respectively. Serum calcium, phosphonium and PTH were assayed and pathological changes observed.
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(1) PTH gene was amplified from human embryonic parathyroid gland tissue, and plasmid pcDNA3.1-PTH-GFP (pcDPG) was constructed by TOPO recombination technique. Digestion, PCR and sequencing were used to identify the positive vectors. (2) pcDPG was transformed into 293 cells by Lipofectamine 2000(TM), fluorescent inverted microscope was used to observe GFP expression, and PTH gene expression was assayed by RT-PCR technique. (3) PTH protein in supernatant was purified and evaluated biological activity. (4) HPT rabbit models were developed and plasmid pcDPG was injected in skeletal muscles, respectively. Serum calcium, phosphonium and PTH were assayed and pathological changes observed.
(1) The findings in digestion and PCR were accorded to anticipation and sequences in report were id</abstract><cop>China</cop><pmid>16677507</pmid><tpages>6</tpages></addata></record> |
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| identifier | ISSN: 0376-2491 |
| ispartof | Zhong hua yi xue za zhi, 2006-01, Vol.86 (4), p.260-265 |
| issn | 0376-2491 |
| language | chi |
| recordid | cdi_proquest_miscellaneous_67945061 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals |
| subjects | Animals Calcium - blood Female Genetic Therapy - methods Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism Humans Hypoparathyroidism - blood Hypoparathyroidism - therapy Mice Mice, Inbred Strains Microscopy, Fluorescence Parathyroid Hormone - blood Parathyroid Hormone - genetics Parathyroid Hormone - metabolism Plasmids - administration & dosage Plasmids - genetics Rabbits Random Allocation Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Reverse Transcriptase Polymerase Chain Reaction Transfection |
| title | pcDPG parathyroid hormone gene therapy of hypoparathyroidism: an experimental study |
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