Construction of a human full-length cDNA bank.
We aimed to construct a full-length cDNA bank from an entire set of human genes and to analyze the function of a protein encoded by each cDNA. To achieve this purpose, a multifunctional phagemid shuttle vector, pKA1, was constructed for preparing a high-quality cDNA library composed of full-length cDNA clones which can be sequenced and expressed in vitro and in mammalian cells without subcloning the cDNA fragment into other vectors. Using this as a vector primer, we have prepared a prototype of the bank composed of full-length cDNAs encoding 236 human proteins whose amino acid sequences are identical or similar to known proteins. Most cDNAs contain a putative cap site sequence, some of which show a pyrimidine-rich conserved sequence exhibiting polymorphism. It was confirmed that the vector permits efficient in vitro translation, expression in mammalian cells and the preparation of nested deletion mutants.
Kato
S
S
Kanagawa Academy of Science and Technology (KAST), Japan.
Sekine
S
S
Oh
S W
SW
Kim
N S
NS
Umezawa
Y
Y
Abe
N
N
Yokoyama-Kobayashi
M
M
Aoki
T
T
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D008322:Mammals; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics*; D015183:Restriction Mapping
D018076:DNA, Complementary; D011506:Proteins