A method for introducing site-specific mutations using oligonucleotide primers and its application to site-saturation mutagenesis

Oligonucleotide primer-directed mutagenesis is a useful molecular biological tool, which is invalubable for the study of the structure/function relationships in proteins and for the creation of mutant proteins possessing modified or novel biological activities. Mutagenesis studies in which a site-saturation approach is employed require a high-efficiency mutagenesis procedure, which will generate a population of mutated molecules containing an even distribution of all possible amino acid changes, or a subset thereof. This article describes such a mutagenesis technique and discusses the adaptations that are necessary to perform sitesaturation mutagenesis.     

γ-Aminobutyric acid (GABA) and other amino acids in tissues of the tick,Amblyomma hebraeum (Acari: Ixodidae) throughout the feeding and reproductive periods

We assayed a variety of tick (Amblyomma hebraeum Koch; Acari, Ixodidae) tissues for a number of amino acids throughout the feeding and early reproductive periods. Our HPLC assay could detect as little as 2–5 pmol per sample of the following: GABA, glycine, serine, glutamine, alanine, taurine, glutamate and aspartate. All of these amino acids could be detected in the salivary gland, synganglion (=total CNS in acarines), haemolymph, Gené's organ, seminal receptacle and ovary. GABA reached high levels in the salivary gland of freshly engorged ticks (685 nmol g^-1) and in the synganglion it exceeded 1000 nmol g^-1 throughout most of the feeding cycle and the first week post-engorgement. GABA also reached a peak titre in the haemolymph of 40 nmol ml^-1. Taurine levels peaked at 1065 nmol g^-1 in the salivary gland from large partially fed ticks. Glutamate and aspartate were likewise found in the salivary gland and synganglion at high concentrations. For most of the amino acids there is insufficient information to correlate these titres (and fluctuations of titres) to neuromodulatory functions. It is possible, however, that the high GABA titre in the salivary gland of engorged ticks is correlated with an augmented level of fluid secretion.Deceased: Department of Pharmacology, University of Alberta, Edmonton, Alberta, Canada, T6G 2E9.     

The relative biological activities of α- and β-ecdysone and their 3 dehydro derivatives in the chromosome puffing assay

The relative biological activity of α- and β-ecdysone and their 3-dehydro derivatives in inducing changes in puffing activity in D. melanogaster salivary glands in vitro is approximately constant. Although the six different puffing assays differed considerably in their sensitivity to ecdysones, no stage-specific or locus-specific differential response to the four compounds was detected.     

Why Do the Karo Batak Prefer Women with Big Feet?

Men may find women with small feet relative to body size more attractive because foot size reliably indexes nubility—i.e., age and parity. I collected judgments of attractiveness in response to drawings of women with varying foot sizes from a sample of 159 Karo Batak respondents from North Sumatra, Indonesia, as part of a collaborative project on foot size and attractiveness. The data revealed a contrarian preference among the Karo Batak for women with big feet. The judgments were compared with the results of an existing cross-cultural study that found a preference for women with small feet in aggregate, but a mix of small- and large-foot preferences in the societies taken individually. Using contingency table analysis, I found that ecology and less exposure to Western media were associated with a preference for women with big feet; patriarchal values were not. The findings suggest that human mating preferences may arise in response to local ecological conditions, and may persist and spread via cultural transmission. This has implications for the concept of universality espoused in some versions of evolutionary psychology.     

Restricted plant species on sub-Antarctic Macquarie and Heard Islands

The recent distributional history of two Macquarie Island vascular plant species, Carex trifida, Poa litorosa, and the Heard Island vascular plant, Ranunculus crassipes is examined. C. trifida is known from only one small population on the north west coast of Macquarie Island. Four populations of P. litorosa were first recorded in the 1980s; we believe however, that it was first observed, but misidentified in the 1950s. R. crassipes was first discovered on Heard Island in the late 1980s. We argue that all three species are indigenous and arrived on their respective islands within the last 200 years by natural processes, most likely from warmer neighbouring islands, where these species have more extensive distributions. There have been small-scale changes in distribution of all species, mainly expansion. Further expansion of all three species is expected as a response to warming climate. Feral rabbit grazing is having a confounding negative influence on populations of P. litorosa.     

Editor’s note

Purinergic Signalling -