[cDNA cloning, sequencing and expression of recombinant human prorelaxin].
In order to accelerate the development of biological recombinant human prorelaxin in our country, direct cloning of human prorelaxin gene from freshly prepared corpus luteum of Chinese woman by RT-PCR method was conducted. Isolated prorelaxin gene contains B, A chains and connective peptide. The amplified products were cloned and confirmed by DNA sequencing using Sanger Dideoxy method. It was subcloned into LKB2, a prokaryotic expression vector offering promoter LacI control, and highly expressed in E. coli. The results suggest that the PCR amplified DNA fragment shared identical sequence with known prorelaxin gene reported abroad. The SDS-PAGE analysis revealed that the expressed protein accounted for about 30% of the total cell protein, its MW was approximately 20 kD and conformed soluble protein.
Ding
C
C
National Laboratory of Molecular Virology and Genetic Engineering, Beijing.
Yang
X
X
Bi
S
S
Fan
L
L
Wen
L
L
Hou
Y
Y
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012065:Relaxin / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D011498:Protein Precursors; D011994:Recombinant Proteins; C038656:prorelaxin; D012065:Relaxin