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The pGEM-T is 3kb in length, and contains the amplicin resistance gene, conferring selection of the plasmid in E. coli, and the ori site which is the bacterial origin of replication. The plasmid has multiple cloning sites as shown below. The coding sequence was inserted by TA cloning. Many E. coli strains are suitable for the propagation of this vector including JM109, DH5α and TOP10.
The coding sequence can be easily obtained by digesting the vector with proper restriction enzyme(s). The coding sequence can also be amplified by PCR with M13 primers, or primer pair SP6 and T7.
SIGLEC6, also known as CD327, belongs to the immunoglobulin superfamily, SIGLEC (sialic acid binding Ig-like lectin) family. SIGLEC6 is a sialic acid recognizing protein expressed at high levels in placenta (cyto- and syncytiotrophoblastic cells) and at lower levels in spleen, peripheral blood leukocytes (predominantly B-cells) and small intestine. SIGLEC6 localizes in various compartments such as membrane fraction, extracellular region and so on. SIGLEC6 may show increasing expression human labor and following childbirth, it has been speculated that this expression helps to slow the tempo of human labor. Interestingly, expression of SIGLEC6 is further upregulated in pre-eclampsia, which appears to be a uniquely human disease.