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M Ku

Publications and source records attributed to M Ku.

8 recordsLinked to original sources
PubMed2000

Developmentally regulated excision of a 28-base-pair sequence from the Paramecium genome requires flanking DNA.

The micronuclear DNA of Paramecium tetraurelia is estimated to contain over 50,000 short DNA elements that are precisely removed during the formation of the transcriptionally active macronucleus. Each internal eliminated sequence (IES) is bounded by 5'-TA-3' dinucleotide repeats, a feature common to some classes of DNA transposons. We have developed an in vivo assay to analyze these highly efficient and precise DNA excision events. The microinjection of a cloned IES into mating cells results in accurately spliced products, and the transformed cells maintain the injected DNA as extrachromosomal molecules. A series of deletions flanking one side of a 28-bp IES were constructed and analyzed with the in vivo assay. Whereas 72 bp of DNA flanking the eliminated region is sufficient for excision, lengths of 31 and 18 bp result in reduced excision and removal of all wild-type sequences adjacent to the TA results in complete failure of excision. In contrast, nucleotide mutations within the middle of the 28-bp IES do not prevent excision. The results are consistent with a functional role for perfect inverted repeats flanking the IES.

Animals↗
PubMed1993

Xenopus axis formation: induction of goosecoid by injected Xwnt-8 and activin mRNAs.

In this study, we compare the effects of three mRNAs-goosecoid, activin and Xwnt-8- that are able to induce partial or complete secondary axes when injected into Xenopus embryos. Xwnt-8 injection produces complete secondary axes including head structures whereas activin and goosecoid injection produce partial secondary axes at high frequency that lack head structures anterior to the auditory vesicle and often lack notochord. Xwnt-8 can activate goosecoid only in the deep marginal zone, i.e., in the region in which this organizer-specific homeobox gene is normally expressed on the dorsal side. Activin B mRNA, however, can turn on goosecoid in all regions of the embryo. We also tested the capacity of these gene products to restore axis formation in embryos in which the cortical rotation was blocked by UV irradiation. Whereas Xwnt-8 gives complete rescue of anterior structures, both goosecoid and activin give partial rescue. Rescued axes including hindbrain structures up to level of the auditory vesicle can be obtained at high frequency even in the absence of notochord structures. The possible functions of Wnt-like and activin-like signals and of the goosecoid homeobox gene, and their order of action in the formation of Spemann's organizer are discussed.

Activins↗
PubMed1993

Xwnt-11: a maternally expressed Xenopus wnt gene.

We have isolated and characterized a novel Xenopus wnt gene, Xwnt-11, whose expression pattern and overexpression phenotype suggest that it may be important for dorsal-ventral axis formation. Xwnt-11 mRNA is present during oogenesis and embryonic development through swimming tadpole stages. Xwnt-11 mRNA is ubiquitous in early oocytes and is localized during mid-oogenesis. By late oocyte stages, Xwnt-11 mRNA is localized to the vegetal cortex, with some mRNA in the vegetal cytoplasm. After egg maturation, Xwnt-11 mRNA is released from the vegetal cortex and is found in the vegetal cytoplasm. This early pattern of Xwnt-11 mRNA localization is similar to another vegetally localized maternal mRNA, Vg1 (D. A. Melton (1987) Nature 328, 80-82). In the late blastula, Xwnt-11 mRNA is found at high levels in the dorsal marginal zone. As gastrulation proceeds, Xwnt-11 mRNA appears in the lateral and ventral marginal zone and, during tadpole stages, it is found in the somites and first branchial arch. Injection of Xwnt-11 mRNA into UV-ventralized embryos can substantially rescue the UV defect by inducing the formation of dorsal tissues. The rescued embryos develop somitic muscle and neural tube; however, they lack notochord and anterior head structures.

Amino Acid Sequence↗
PubMed1989

The bovine pericardial xenograft: III. Effect of uniaxial and sequential biaxial stress during fixation on the tensile viscoelastic properties of bovine pericardium.

Our previous two articles have shown that glutaraldehyde-fixed bovine pericardium is nearly isotropic, whether fixed without constraints, with tethering, or with pressure. In this study, we have used uniaxial stress during fixation to produce bovine pericardial material with marked tensile anisotropy. Rectangular and cruciate pericardial samples have been mechanically examined after one of four treatments: (i) fixation under 88-kPa uniaxial stress, (ii) fixation under 176-kPa uniaxial stress, (iii) 3 h of 176-kPa uniaxial stress in saline followed by 24-h fixation under the same stress, (iv) fixation under 176-kPa uniaxial stress followed by a second fixation under 176-kPa stress in a direction normal to the first. Strips of material were cut at 0 degree, 30 degrees, 60 degrees, and 90 degrees to direction of the initial stress, and tested for response to cyclic loading, stress relaxation, plastic deformation, and fracture properties. Fixation under uniaxial stress produced anistropy similar to that seen in porcine aortic valve leaflets; however, the overall extensibility of the material depended on the applied stress and the aspect ratio of the stressed sample. While loading in saline produced no change, the sequential biaxial stressing produced a reduction in anisotropy, suggesting exposure of additional crosslinking sites. Uniaxial stress during fixation may be a useful method for construction of anisotropic heart valve leaflets.

Animals↗