Angiogenin Expression in Plants
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example 1
Cloning of the Bovine Angiogenin Gene
[0178]The Bos taurus (cow) angiogenin, ribonuclease, RNase A family, 5 (ANG), mRNA sequence is available from the National Centre for Biotechnology Information (NCBI), accession number AM—0011078144. The predicted open reading frame (ORF) contains 444 base pairs (bp) (FIG. 1) encoding a 148 amino acid (aa) (FIG. 2) sequence. Using the SignalP 3.0 server to predict the presence and location of signal peptide cleavage sites in amino acid sequences a 24 aa (72 bp) signal peptide sequence was identified (FIGS. 1 and 2).
[0179]The angiogenin protein sequence has been analysed by comparison to a database of known allergens, the Food Allergy and Resource Research Program at the University of Nebraska allergen protein database (FARRP Allergen Online version 7.0). A BLASTp for every 80 amino acid peptides contained within the protein was searched against the FAARP Allergen Online database. None of the amino acid peptides contained 35% or higher identity to...
example 2
Production of Fusion Proteins for Greater Accumulation, Enhanced Action, or Improved Extraction, of Angiogenin
[0183]It is possible to create fusion proteins of angiogenin with mediators or modulators of its activity to assist in the improvement of protein accumulation, enhancement of protein action, or for effective extraction of the protein.
Fusion Proteins for Enhancing the Action of Angiogenin
[0184]Yeast two-hybrid technology has identified potential ANG-interacting molecules (Goa and Xu, 2008) such as alpha-actin 2 (ACTN-2) (Hu et al., 2005), regulatory proteins such as follistatin (FS) (Goa et al., 2007) and extracellular matrix proteins such as fibulin-1 (Zhang et al., 2008). It is hypothesised that through interacting with ACTN-2, ANG may regulate the movement or the cytokinesis of the cells, follistatin may act as a regulator on angiogenin's actions and interaction between ANG and fibulins may facilitate cell adhesion.
[0185]Follistatin is known to have a role in muscle growth...
example 3
Identification of Promoter Sequences for Targeted Expression of Angiogenin
[0188]Promoters with tissue-specificity are required to drive expression of transgenes in crops to target accumulation in particular tissues / organs and to avoid unwanted expression elsewhere. Therefore highly expressing but yet tightly controlled promoters are desirable.
Tissue Specific or Regulated Promoters
[0189]The choice of promoters affects transgene expression concentration, as well as developmental, tissue and cell specificity. Examples of different promoters to drive transgene expression for different objectives are presented in Table 1.
TABLE 1Examples of different promoters to drive transgene expression.Targeted expressionGene promoterOrganismReferenceConstitutiveConstitutiveUbiquitin, UbiZea mays (maize)Christensenet al. (1992)CaMV35S2CauliflowerKay et al.mosaic virus(1987)Polyubiquitin, RUBQ2Oryza sativa (rice)Liu et al.(2003)Actin 1, OsAct1Oryza sativa (rice)McElroyet al. (1990)Tissue SpecificTuber ...
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