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Molecular cloning of DNA sequences coding for mouse embryonic globins
Acta Biologica Et Medica Germanica
|January 1, 1981
Summary
Researchers synthesized complementary DNAs (cDNAs) for mouse embryonic globins. They cloned these cDNAs into plasmids, successfully isolating and characterizing clones coding for specific embryonic globin chains.
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- Mouse embryonic erythroid cells produce four distinct embryonic globins.
- Understanding the genetic basis of these globins is crucial for studying early hematopoiesis.
Purpose of the Study:
- To clone and characterize complementary DNAs (cDNAs) encoding mouse embryonic globins.
- To identify specific embryonic globin chains represented by the cloned cDNAs.
Main Methods:
- Synthesis of double-stranded cDNAs from purified globin messenger RNAs.
- Ligation of cDNAs into a pBR322 plasmid vector and transformation of E. coli.
- Screening of bacterial clones via hybridization and characterization using restriction mapping and hybrid translation.
Main Results:
- Isolation of recombinant plasmids containing embryonic globin cDNAs.
- Characterization revealed varying sizes of inserted sequences and loss of tetracycline resistance genes in some plasmids.
- Hybrid translation identified plasmid no. 2 as coding for an alpha-like chain (x), while plasmids no. 16 and 54 code for beta-like chains (y or z).
Conclusions:
- Successful cloning and initial characterization of cDNAs for mouse embryonic globins.
- Demonstrated the ability to differentiate between alpha-like and beta-like embryonic globin chains using molecular techniques.