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Manipulation of tyrosine decarboxyla...
~
Lee, Jintae.
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Manipulation of tyrosine decarboxylase expression in California poppy cells.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Manipulation of tyrosine decarboxylase expression in California poppy cells./
作者:
Lee, Jintae.
面頁冊數:
98 p.
附註:
Source: Dissertation Abstracts International, Volume: 65-06, Section: B, page: 3038.
Contained By:
Dissertation Abstracts International65-06B.
標題:
Engineering, Chemical. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3134854
ISBN:
0496820799
Manipulation of tyrosine decarboxylase expression in California poppy cells.
Lee, Jintae.
Manipulation of tyrosine decarboxylase expression in California poppy cells.
- 98 p.
Source: Dissertation Abstracts International, Volume: 65-06, Section: B, page: 3038.
Thesis (Ph.D.)--Rutgers The State University of New Jersey - New Brunswick, 2004.
The production of benzophenanthridine alkaloids (BPAs) from suspension cell cultures of Eschscholzia californica (California poppy) has been extensively studied with an emphasis on strategies to overcome low productivity. The main objective of this work is to investigate how changes in expression of genes coding for a potential regulatory enzyme in the BPA pathway effect final production yields, selectivities, and productivities.
ISBN: 0496820799Subjects--Topical Terms:
1018531
Engineering, Chemical.
Manipulation of tyrosine decarboxylase expression in California poppy cells.
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The production of benzophenanthridine alkaloids (BPAs) from suspension cell cultures of Eschscholzia californica (California poppy) has been extensively studied with an emphasis on strategies to overcome low productivity. The main objective of this work is to investigate how changes in expression of genes coding for a potential regulatory enzyme in the BPA pathway effect final production yields, selectivities, and productivities.
520
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We have established a rapid and stable genetic transformation protocol suited to California poppy using disarmed Agrobacterium tumefaciens as a vector carrying a green fluorescent protein (GFP) under the control of the constitutive cauliflower mosaic virus (CaMV) 35S promoter. The transformed poppy cells have expressed high levels of GFP and maintained this phenotypic characteristic for over 4 years.
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Transformation of poppy cells was also carried out for the purpose of introducing elevated enzyme activity into the BPA pathway. Tyrosine decarboxylase (TYDC) converts L-tyrosine into tyramine as the first committed step in the biosynthesis of BPAs from poppy-fumaria species. Transformed California poppy cells expressing a fusion protein of GFP and opium poppy TYDC under the control of CaMV 35S promoter were established. The transformed cells constitutively express the fusion protein GFP::TYDC, which was localized largely around the nucleus and the plasma membrane. Overexpression of GFP::TYDC did not adversely affect cell growth. The transformed cells displayed more than a 10-fold higher level of tyramine, a direct TYDC enzyme product, compared to controls. The accumulation of BPAs was correlated with the observed tyramine.
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Finally, we sought to transform cells with the purpose of controlling gene expression in response to nontoxic, simple environmental cues. A fusion gene of the Arabidopsis methyl jasmonate (MeJA)/elicitor-inducible PDF1.2 promoter and GFP was established and transformed in California poppy cells to demonstrate that the PDF1.2 promoter acts as an inducible promoter. Expression of GFP is both dose-dependent and time-dependent in response to MeJA or yeast elicitor. GFP expression is more rapidly induced with yeast elicitor compared to MeJA. Compared to the basal expression level, the level of GFP expression was increased more than 10-fold within 9 hours after treatment of yeast elicitor.
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http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3134854
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