SOMA: A Single Oligonucleotide Mutagenesis and Cloning Approach

Modern biology research requires simple techniques for efficient and restriction site-independent modification of genetic material. Classical cloning and mutagenesis strategies are limited by their dependency on restriction sites and the use of complementary primer pairs. Here, we describe the Single Oligonucleotide Mutagenesis and Cloning Approach (SOMA) that is independent of restriction sites and only requires a single mutagenic oligonucleotide to modify a plasmid. We demonstrate the broad application spectrum of SOMA with three examples. First, we present a novel plasmid that in a standardized and rapid fashion can be used as a template for SOMA to generate GFP-reporters. We successfully use such a reporter to assess the in vivo knock-down quality of morpholinos in Xenopus laevis embryos. In a second example, we show how to use a SOMA-based protocol for restriction-site independent cloning to generate chimeric proteins by domain swapping between the two human hRMD5a and hRMD5b isoforms. Last, we show that SOMA simplifies the generation of randomized single-site mutagenized gene libraries. As an example we random-mutagenize a single codon affecting the catalytic activity of the yeast Ssy5 endoprotease and identify a spectrum of tolerated and non-tolerated substitutions. Thus, SOMA represents a highly efficient alternative to classical cloning and mutagenesis strategies.     

Chromosomes and reproductive behaviour of Finnish Taraxacum agamospecies

Material consisting of 157 accessions of 69 Finnish Taraxacum agamospecies, representing 7 sections, was grown in an experimental field for comparison of certain botanical and nutritional characteristics. This paper gives the collecting data as well as cytological and reproductive characteristics of the material. The largest section Taraxacum (Vulgariu ) was represented by 60 agamospecies, 59 of them triploid (2n = 24) and one tetraploid (T. penicilliforme , 2n = 32). The tetraploid and 37 triploids were chromosome counted for the first time. The diploid sexual species T. obtusilobum from Sweden was included for comparison. Other sections consisted of 3 triploids and 6 tetraploids, 3 tetraploid species of the section Erythrosperma being chromosome counted for the first time. Seedlings with deviating numbers were occasionally found. A banding technique of the chromosomes was developed but it did not allow the exact identification of other than the satellite chromosomes. Species-specific features were found in the flowering phenology and in several pollen characteristics (stainability, mean and variability of the size). There were indications of genetic variation within certain agamospecies in pollen characteristics. Isolated and emasculated (mostly only isolated) flower heads gave mostly a full seed set, which indicates obligate, autonomous agamospermy. Lower seed sets of T. adami, T. dahlstedtii and T. hamatiforme in isolation may be a sign of facultative apomixis in these species, requiring further study.     

Site factors are more important than management for indicator species in semi-natural grasslands in southern Sweden

Plant Ecology - Management of semi-natural grasslands is essential to retain the characteristic diversity of flora and fauna found in these habitats. To maintain, restore or recreate favourable...     

Identification of copy number variants in children and adolescents with autism spectrum disorder: a study from Turkey

Molecular Biology Reports - Copy number variants (CNVs) play a key role in the etiology of autism spectrum disorder (ASD). Therefore, recent guidelines recommend chromosomal microarrays (CMAs) as...     

Two Metabolic Fuels,Glucose and Lactate,Differentially Modulate Exocytotic Glutamate Release from Cultured Astrocytes

Neurochemical Research - Astrocytes have a prominent role in metabolic homeostasis of the brain and can signal to adjacent neurons by releasing glutamate via a process of regulated exocytosis....