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Secretion of active recombinant phyt...

Li, Jia.

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Secretion of active recombinant phytase from stably transformed soybean cells.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Secretion of active recombinant phytase from stably transformed soybean cells./
Author:	Li, Jia.
Description:	140 p.
Notes:	Source: Dissertation Abstracts International, Volume: 57-02, Section: B, page: 0893.
Contained By:	Dissertation Abstracts International57-02B.
Subject:	Biology, Molecular. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9618981

Secretion of active recombinant phytase from stably transformed soybean cells.
Li, Jia.

Secretion of active recombinant phytase from stably transformed soybean cells. - 140 p.

Source: Dissertation Abstracts International, Volume: 57-02, Section: B, page: 0893.

Thesis (Ph.D.)--Virginia Polytechnic Institute and State University, 1995.

The objective of this research was to express a fungal phytase gene in transgenic soybean cells to to study the potential for improving phosphorus utilization in soybean meal. A simple and inexpensive particle inflow gene gun was constructed and bombardment was optimized as assayed by Subjects--Topical Terms:

1017719
Biology, Molecular.

Secretion of active recombinant phytase from stably transformed soybean cells.
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008 130515s1995 ||||||||||||||||| ||eng d
035 $a (UMI)AAI9618981
035 $a AAI9618981
040 $a UMI $c UMI
100 1 $a Li, Jia. $3 1250927
245 1 0 $a Secretion of active recombinant phytase from stably transformed soybean cells.
300 $a 140 p.
500 $a Source: Dissertation Abstracts International, Volume: 57-02, Section: B, page: 0893.
500 $a Chairperson: Elizabeth A. Grabau.
502 $a Thesis (Ph.D.)--Virginia Polytechnic Institute and State University, 1995.
520 $a The objective of this research was to express a fungal phytase gene in transgenic soybean cells to to study the potential for improving phosphorus utilization in soybean meal. A simple and inexpensive particle inflow gene gun was constructed and bombardment was optimized as assayed by $\ beta $- glucuronidase reporter gene expression. A somatic embryogenesis approach was used for soybean regeneration from culture. The efficiencies of embryo induction and embryo conversion to form roots and shoots were compared in commercial soybean cultivars to identify optimal cultivars for recovery of transgenic plants.
520 $a To study the expression of a recombinant fungal phytase gene (phyA from Aspergillus niger), four expression vectors were constructed in soybean transformation vectors. PhyA was placed under the control of either a constitutive cauliflower mosaic virus 35S promoter or a soybean seed specific $\ beta $- conglycinin promoter, each with or without a patatin endoplasmic reticulum (ER) signal sequence. All four vectors were sequenced and introduced into 'Williams 82' suspension culture cells by particle bombardment. Stably transformed cell lines were selected and tested for stable integration by Southern analysis. The presence of the phytase protein product was detected by immunoblotting. Activity of recombinant phytase was characterized by enzyme assay. Cell lines containing the phyA gene under control of the CaMV 35S promoter and ER signal sequence secreted active phytase into the culture medium. The pH and temperature optima were determined for recombinant phytase.
520 $a Two constructs containing the ER signal sequence (both the constitutive and seed specific promoters) were used to bombard regenerable soybean somatic embryos. Hygromycin resistant embryos were recovered after six weeks of selection. Embryos were transferred to development medium for regeneration. Several embryos formed roots and shoots but have not yet progressed to fertile regenerated plants.
590 $a School code: 0247.
650 4 $a Biology, Molecular. $3 1017719
650 4 $a Biology, Cell. $3 1017686
650 4 $a Biology, Plant Physiology. $3 1017865
690 $a 0307
690 $a 0379
690 $a 0817
710 2 $a Virginia Polytechnic Institute and State University. $3 1017496
773 0 $t Dissertation Abstracts International $g 57-02B.
790 1 0 $a Grabau, Elizabeth A., $e advisor
790 $a 0247
791 $a Ph.D.
792 $a 1995
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9618981