Isolation and characterization of the messenger RNA and the gene coding for a proline-rich zein from corn endosperm
Gamma-zein, a proline-rich protein from corn endosperm, was investigated at the molecular level. Immunological and electrophoretic data indicated that gamma-zein was deposited into protein bodies in corn endosperm. Both isolated polysomes and poly(A)/sup +/ mRNA were found to direct into vitro synthesis of gamma-zein in a wheat germ system. In vitro synthesized gamma-zein was immunoprecipitated from the total in vitro translation products. A cDNA expression library was constructed by reverse transcription of total poly(A)/sup +/ mRNA using pUC8 plasmid as vector and E. coli strain DH1 as host. The library was screened for the expression of gamma-zein and alpha-zein by specific antibodies. The library was also screened with /sup 32/P-labeled gamma-zein and alpha-zein cDNA probes. The results indicated that gamma-zein and its fragments were readily expressed in E. coli while alpha-zein was not. Seven independently selected clones, six of which were selected by antibody and one by a cDNA probe, were sequenced. A comparison of sequence information from seven clones revealed that their overlapping regions were identical. This suggests that gamma-zein is encoded by a single gene. This finding is in conflict with what was expected on the basis of extensive charge heterogeneity of gamma-zein in isoelectric focusing. Individual bands cut from an IEF gel were rerun and shown to give several bands suggesting that the charge heterogeneity of gamma-zein may be an artifact. Sequence information of gamma-zein indicated that the gene encodes a mature protein whose primary structure includes 204 amino acids and has a molecular weight of 21,824 daltons.
- Research Organization:
- Virginia Polytechnic Inst. and State Univ., Blacksburg (USA)
- OSTI ID:
- 5008986
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
Similar Records
Heterogeneity and expression of the zein storage protein gene family of maize
The molecular biology and biochemistry of rice endosperm. alpha. -globulin
Related Subjects
ENDOSPERM
MOLECULAR BIOLOGY
PROTEINS
BIOSYNTHESIS
AMINO ACID SEQUENCE
GENES
MAIZE
MESSENGER-RNA
PHOSPHORUS 32
PROLINE
AMINES
AMINO ACIDS
AZOLES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CEREALS
DAYS LIVING RADIOISOTOPES
GRASS
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
ISOTOPES
LIGHT NUCLEI
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS ISOTOPES
PLANT TISSUES
PLANTS
PYRROLES
PYRROLIDINES
RADIOISOTOPES
RNA
SYNTHESIS
TISSUES
550201* - Biochemistry- Tracer Techniques
550401 - Genetics- Tracer Techniques