In vitro site-directed mutagenesis with synthetic DNA oligonucleotides yields unexpected deletions and insertions at high frequency.

We have used in vitro site-directed mutagenesis with synthetic DNA oligonucleotides to introduce single nucleotide mutations in yeast mtDNA. In addition to the expected DNA alterations we also recovered with high frequency mutants with large deletions and insertions which arose through interaction with the synthetic DNA fragment. Characterization of a number of these by DNA sequence analysis has permitted reconstruction of the mutagenic events. In all cases, the DNA fragment had base paired with non-adjacent DNA sequences sometimes more than 1000 nucleotides apart from each other on the target strand. The products of such interactions cannot be avoided due to the non-stringent annealing conditions during complementary DNA strand synthesis. However, deliberate mispairing can be directed precisely, as shown by our ability to specifically delete the 1143-bp intron from the yeast mitochondrial gene coding for large ribosomal RNA with a synthetic DNA fragment consisting of the sequence of the exon borders flanking the intron.     

Synthesis of short RNA fragments by the benzotriazolyl phosphotriester approach

Analytical and experimental evidence will be presented to show that addition of a slight excess of 2-chlorophenyl-0,0-bis[1-benzotriazolyl]phosphate to a properly 2',5'-protected ribonucleoside gives an intermediate 3'-phosphotriester, which, after addition of a 2'-protected ribonucleoside, affords a dimer containing solely a 3'-5'-internucleotide linkage. The above phosphorylation procedure has been used in the synthesis of the pentamer UpApCpGpC.     

In vitro incorporation of 2′-deoxyadenosine and 3′-deoxyadenosine into yeast tRNAPhe using tRNA nucleotidyl transferase,and properties of tRNAPhe-C-C-2′dA and tRNAPhe-C-C-3′dA

2′-Deoxyadenosine and 3′-deoxyadenosine (cordycepin) can be incorporated into the 3′-terminal position of tRNA^Phe by tRNA nucleotidyl transferase. tRNA^Phe-C-C-2′dA and tRNA^Phe-C-C-3′dA, missing the cis-diol group at the 3′-terminal end are resistant to periodate oxidation and are not able to form borate complexes. In aminoacylation experiments only the tRNA^Phe-C-C-3′dA proved to be chargeable.     

Zur Frage der Verteilung des Rh(D)-Faktors bei erstgeborenen und nachgeborenen Kindern

Summary The frequency of the occurrence of the Rh(D)-factor was compared in 1185 first-born babies with that among 2246 children from 2nd and later pregnancies. Among the first-born children 14,2% were Rh(D)-negative, and among the later-born 14,7% were Rh(D)-negative. In children with the Blood group O the distribution of the Rh(D) bloodgroup was the same in both first-born and later-born children. Among the Rh(D)-positive children there seem to be less boys.     

Beitrag zur Kenntnis der österreichischen Sippen desValeriana officinalis-Aggregats und ihrer Chromosomenzahlen

Zusammenfassung Die vonWalther im wesentlichen für Deutschland ausgearbeitete Gliederung vonValeriana officinalis agg. ist in ihren Grundzügen auch in Österreich anwendbar. Die Chromosomenzahlen der Sippen sind in diesem Land die gleichen wie auch in anderen Teilen Europas.Valeriana officinalis L. s. str. — die in Österreich häufigste Art — ist mit 2n=14 diploid,V. collina Wallr. mit 2n=28 tetraploid. Ob die beiden in Österreich weniger häufigen, einander sehr ähnlichen oktoploiden Sippen mit 2n=56 —V. sambucifolia Mikan fil. undV. procurrens Wallr. — wirklich Artrang verdienen, ist zu diskutieren.V. procurrens wächst in Österreich unter anderem in den Nordtiroler Alpen.

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Summary The subdivision ofValeriana officinalis agg. established byWalther especially for Germany is found to be practicable in its main features in Austria too. The chromosome numbers of the taxa are the same in Austria as in other parts of Europe.Valeriana officinalis L. s. str. is a diploid (2n=14),V. collina Wallr. is a tetraploid species (2n=28). As to the nearly related octoploid taxaV. sambucifolia Mikan fil. andV. procurrens Wallr. (2n=56) it is still to be discussed, if they should be recognized as species or not.

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Herrn Professor Dr. L.Geitler zum 70. Geburtstag gewidmet.