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M L Hoffman

Publications and source records attributed to M L Hoffman.

12 recordsLinked to original sources

Structural and functional analysis of chicken U4 small nuclear RNA genes.

Two distinct chicken U4 RNA genes have been cloned and characterized. They are closely linked within 465 base pairs of each other and have the same transcriptional orientation. The downstream U4 homology is a true gene, based on the criteria that it is colinear with chicken U4B RNA and is expressed when injected into Xenopus laevis oocytes. The upstream U4 homology, however, contains seven base substitutions relative to U4B RNA. This sequence may be a nonexpressed pseudogene, but the pattern of base substitutions suggests that it more probably encodes a variant yet functional U4 RNA product not yet characterized at the RNA level. In support of this, the two U4 genes have regions of homology with each other in their 5'-flanking DNA at two positions known to be essential for the efficient expression of vertebrate U1 and U2 small nuclear RNA genes. In the case of U1 and U2 RNA genes, the more distal region (located near position-200 with respect to the RNA cap site) is known to function as a transcriptional enhancer. Although this region is highly conserved in overall structure and sequence among U1 and U2 RNA genes, it is much less conserved in the chicken U4 RNA genes reported here. Interestingly, short sequence elements present in the -200 region of the U4 RNA genes are inverted (i.e., on the complementary strand) relative to their usual orientation upstream of U1 and U2 RNA genes. Thus, the -200 region of the U4 RNA genes may represent a natural evolutionary occurrence of an enhancer sequence inversion.

Animals↗

Prevention of monkey sperm penetration of zona-free hamster ova by sperm antibody obtained from vasectomized cynomolgus monkeys.

Incubation of antisperm sera from vasectomized cynomolgus monkeys (Macaca fascicularis) with monkey sperm caused a fourfold reduction of sperm attachment to and penetration of zona-free Golden hamster ova. The attachment of sperm to hamster ova was reduced from 64% to 11%, and the penetration of ova was reduced from 20% to 5%. Sperm antibodies block sperm attachment to the vitelline membrane, thus preventing ovum penetration. This blockage may be one of the reasons for low fertility rates observed following reanastomosis of the vas deferens in those vasectomized males that show high levels of circulating antisperm antibody.

Animals↗

Sex differences in moral internalization and values.

The subjects were fifth- and seventh-grade white middle-class children and their parents. The major moral internalization indices pertain to internal moral judgment, guilt intensity, and fear of punishment. The findings support the prevalent view that consideration for others is more salient in females. They also suggest, with considerable consistency (especially in adults) that moral transgressions are more likely to be associated with guilt in females and fear in males. No sex differences in internal moral judgment were obtained. Evidence was presented suggesting that the differences in children may be due partly to different discipline and affection patterns. It was also suggested that the results for adults as well as children might be explained by differential sex-role socialization as well as by increasing pressures on males over the life cycle to achieve and succeed, which may often conflict with concerns about the welfare of others.

Achievement↗

Metabolic engineering of propanediol pathways.

Microbial fermentation is an important technology for the conversion of renewable resources to chemicals. In this paper, we describe the application of metabolic engineering for the development of two new fermentation processes: the microbial conversion of sugars to 1,3-propanediol (1,3-PD) and 1,2-propanediol (1,2-PD). A variety of naturally occurring organisms ferment glycerol to 1,3-PD, but no natural organisms ferment sugars directly to 1,3-PD. We first describe the fed-batch fermentation of glycerol to 1,3-PD by Klebsiella pneumoniae. We then present various approaches for the conversion of sugars to 1,3-PD, including mixed-culture fermentation, cofermentation of glycerol and glucose, and metabolic engineering of a "sugars to 1,3-PD" pathway in a single organism. Initial results are reported for the expression of genes from the K. pneumoniae 1,3-PD pathway in Saccharomyces cerevisiae. The best naturally occurring organism for the fermentation of sugars to 1,2-PD is Thermoanaerobacterium thermosaccharolyticum. We describe the fermentation of several different sugars to 1,2-PD by this organism in batch and continuous culture. We report that Escherichia coli strains engineered to express either aldose reductase or glycerol dehydrogenase convert glucose to (R)-1,2-PD. We then analyze the ultimate potential of fermentation processes for the production of propanediols. Linear optimization studies indicate that, under aerobic conditions, propanediol yields that approach the theoretical maximum are possible and CO2 is the primary coproduct. Without the need to produce acetate, final product titers in the range of 100 g/L should be possible; the high titers and low coproduct levels should make product recovery and purification straightforward. The examples given in this paper illustrate the importance of metabolic engineering for fermentation process development in general.

Biotechnology↗