Representation of cloned genomic sequences in two sequencing vectors: correlation of DNA sequence and subclone distribution.
نویسندگان
چکیده
Representation of subcloned Caenorhabditis elegans and human DNA sequences in both M13 and pUC sequencing vectors was determined in the context of large scale genomic sequencing. In many cases, regions of subclone under-representation correlated with the occurrence of repeat sequences, and in some cases the under-representation was orientation specific. Factors which affected subclone representation included the nature and complexity of the repeat sequence, as well as the length of the repeat region. In some but not all cases, notable differences between the M13 and pUC subclone distributions existed. However, in all regions lacking one type of subclone (either M13 or pUC), an alternate subclone was identified in at least one orientation. This suggests that complementary use of M13 and pUC subclones would provide the most comprehensive subclone coverage of a given genomic sequence.
منابع مشابه
In silico cloning and bioinformatics study of Brucella melitensis Omp31 antigen in different mammalian expression vectors
Brucella melitensis, as a pathogenic gram-negative intracellular bacterium, causes brucellosis in animals and humans. According to literature, the B. melitensis outer membrane protein 31 (Omp31) is considered as an important vaccine candidate against brucellosis. The aim of the current study was to compare three different expression constructs containing B. melitensis Omp31 antigen using bioinf...
متن کاملCloning and sequencing of Toxoplasma gondii major surface antigen (SAG1) gene
Genetic typing methods of T. gondii strains have been extensively perfected in recent years. From a technical point of view, many tools usable for genetic studied on single-copy loci have been used: RFLP, PCR-RFLP, sequencing, RAPD-PCR and isoenzyme analysis. We described the cloning and sequence analysis of the gene which encodes the major surface antigen (SAG1 or P30) of T. gondii. SAG1 is ...
متن کاملIsolation and expression of the lysis genes of Actinomyces naeslundii phage Av-1.
Like most gram-positive oral bacteria, Actinomyces naeslundii is resistant to salivary lysozyme and to most other lytic enzymes. We are interested in studying the lysins of phages of this important oral bacterium as potential diagnostic and therapeutic agents. To identify the Actinomyces phage genes encoding these species-specific enzymes in Escherichia coli, we constructed a new cloning vector...
متن کاملConstruction of expressing vectors including melanoma differentiation-associated gene-7 (mda-7) fused with the RGD sequences for better tumor targeting
Objective(s): Up to now, many researches have been performed to improve the antitumoral effect of melanoma differentiation-associated gene-7 (mda-7) protein. The purpose of our research was to construct 3 expression vectors producing mda-7 in fusion with RGD (Arginine-Glycine-Aspartic acid) peptide and evaluate their expression. Materials and Methods: mda-7 gene with two different RGD sequ...
متن کاملResistance of Cloned 1F5 Chimeric Anti-CD20 Antibody Heavy-Chain Gene to DNA Polymerase due to a Predicted Hairpin Structure
Background: Formation of secondary structure such as DNA hairpins or loops may influence molecular genetics methods and PCR based approaches necessary for genetic engineering, in addition to gene regulation. Materials and Methods: A polymerase chain reaction with splice overlap extension (SOE-PCR) was used to create fully synthetic 1F5 chimeric anti-CD20 heavy- and light-chain genes. The chi...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Nucleic acids research
دوره 25 15 شماره
صفحات -
تاریخ انتشار 1997