I. No gene transcription occurs in fertilized eggs of the amphibian Xenopus until the mid-blastula transition, (approximately 4000 cells). Therefore, all of the appropriate gene products necessary for early development must be stored in the oocyte or egg prior to fertilization. In order to understand the molecular embryology of early amphibian development, we must obtain genes whose products are expressed only in oocytes. We have been working on an oocyte specific gene product called OAX RNA for oocyte activated in Xenopus. This RNA is 181 nucleotides long, present approximately in 10,000 copies/oocyte and is in a cytoplasmic complex of approximately 50S size. This RNA is not found in adult somatic tissue. II. The Aspergillus toxin alpha-sarcin produces a precise cut near the 3'-end of 28S ribosomal RNA in vitro only if the ribosomes are pre-treated with puromycin and EDTA. Alpha-sarcin can also behave as a general nuclease in vitro under appropriate conditions. In order to investigate alpha-sarcin's in vivo activity, we injected it into living Xenopus oocytes and analyzed the resulting RNA. III. The human and rodent LINE-1 or L1 family of repetitive DNA is present more than 10,000 times per haploid genome. The L1 family has long open reading frames and transcripts present in various types of cells. Using a probe from the 3'-end of a mouse L1 gene, we looked for the presence of RNA transcripts in Xenopus oocytes, embryonic and adult tissues.

Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
U.S. National Inst Diabetes/Digst/Kidney
Department
Type
DUNS #
City
State
Country
United States
Zip Code