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Miniature pig is an attractive animal for a wide range of research fields, such as medicine and pharmacology, because of its small size, the possibility of breeding it under minimum environmental controls and the physiology that is potentially similar to that of human. Although transgenic technology is useful for the analysis of gene function and for the development of model animals for various diseases, there have not yet been any reports on producing transgenic miniature pig. This study is the first successful report concerning the production of transgenic miniature pig by pronuclear microinjection. The huntingtin gene cloned from miniature pig, which is a homologue of candidate gene for Huntington's disease, connected with rat neuron-specific enolase promoter region, was injected into a pronucleus of fertilized eggs with micromanipulator. The eggs were transferred into the oviduct of recipient miniature pigs, whose estrus cycles were previously synchronized with a progesterone analogue. A total of 402 injected eggs from 171 donors were transferred to 23 synchronized recipients. Sixteen of them maintained pregnancy and delivered 65 young, and one resulted in abortion. Five of the 68 offspring (three of which were aborted) were determined to have transgene by PCR and Southern analysis. The overall rate of transgenic production was 1.24% (transgenic/injected eggs). This study provides the first success and useful information regarding production of transgenic miniature pig for biomedical research.

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of cDNA for human tumor necrosis Factor d (TNF-j?) have a discrepancy within the coding region as well as cxon 1. To resolve these discrepancies WC bavc rc-isolated TNF-p cDNA from the human B ccl1 lymphoblasloid ccl1 line. RPM1 1788, and determined its DNA scqucncc. Results indicate that amino acid 26 is threoninc (Thr) insicad of aspanginc (Asn). In contrast to published sequences, the fiequcncc ol' the exon I region corresponded to the genomic suqucncc of TNF;B. From our studies WC conclude that the TNF-/3 gene of the human B cell lympboblastoid cell line, RPM1 1788. is homologous with respect to the TNF-P gene.

Fifty-one cases of human renal cell carcinoma were investigated for immunohistochemical localization of ras oncogenes and their products. The immunohistochemical results were evaluated comparatively with the histological grade, stage and progression of the tumor. The ras p21 oncogene product was positive mainly in the plasma membrane of the renal cell carcinomas. The tissues from normal kidneys were limitedly stained in the cytoplasm of the proximal convoluted tubule cells. Five of 28 (17.9%) frozen specimens of renal cell carcinoma revealed the positive staining for ras mRNA by means of in situ hybridization as well as the strongly positive staining for ras p21 gene products. These 5 cases were all grade 2 and pT2b in TNM classification. However, there was no point mutation on exon 1(corresponding to codons 12 and 13) and exon 2 (corresponding to codon 61), where point mutations are reported in several human tumors. Positive staining cases for ras p21 (survival rate of 81.2% at 5 years) showed the tendency to decrease the survival rate compared to the ras p21 negative staining cases. There was a significant correlation of ras p21 positive staining and the pathological extent of the primary tumor (pT) (p<0.05).Our immunohistochemical study revealed that the expression of ras p21 is one of the parameters related to the prognosis of renal cell carcinomas.

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