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IPC

C12N15/10(2006-01-01)

C12N1/21(2006-01-01)

(21) (22)

Application

2013141777/10, 2013-09-11

(24)

Start date

2013-09-11

(22)

Actual filing date

2013-09-11

(45)

Published

2015-01-10

(72)

Inventor

Balabanova Larisa Anatol'EvnaKoval'Chuk Svetlana NikolaevnaGolotin Vasilij AleksandrovichBulgakov Aleksandr AleksandrovichRasskazov Valerij Aleksandrovich

(73)

Holder

Federal'Noe Gosudarstvennoe Bjudzhetnoe Uchrezhdenie Nauki Tikhookeanskij Institut Bioorganicheskoj Khimii Im. G.B. Eljakova Dal'Nevostochnogo Otdelenija Rossijskoj Akademii Nauk Dvo Ran)

RECOMBINANT PLASMID DNA pET40CmAP/MBL-T CODING HYBRID BIFUNCTIONAL POLYPEPTIDE CmAP/MBL-T WITH PROPERTIES OF HIGH-ACTIVITY ALKALINE PHOSPHATASE CmAP AND MANNAN-BINDING LECTIN OF C-TYPE MBL-T, RECOMBINANT STRAIN E coli Rosetta(DE3)/pET40CmAP/MBL-T - PRODUCER OF HYBRID BIFUNCTIONAL POLYPEPTIDE CmAP/MBL-T, AND METHOD FOR ITS OBTAINING Russian patent published in 2015 - IPC C12N15/10 C12N1/21

Abstract RU 2538169 C1

FIELD: biotechnologies.

SUBSTANCE: invention represents plasmid pET40CmAP/MBL-T determining a synthesis of hybrid bifunctional polypeptide CmAP/MBL-T with properties of high-activity alkaline phosphatase of sea bacterium Cobetia marina (CmAP) and mannan-binding lectin of C-type of far-eastern trepang Apostichopus japonicus (MBL-T). The plasmid includes NcoI/SalI - a fragment of plasmid pET-40b(+) (Novagen) and a DNA fragment with size of 2034 pairs of bases, which is a chimeric gene coding structural genes of alkaline phosphatase CmAP and mannan-binding lectin of C-type MBL-T, which are connected to each other with a polynucleotide coding the amino acid sequence of linker (G)₄S(G)₄S(G)₄SEL. The invention describes recombinant strain E.coli Rosetta(DE3)/pET40CmAP/MBL-T transformed by the above plasmid - a producer of hybrid bifunctional protein CmAP/MBL-T. The invention also proposes a method for obtaining hybrid bifunctional protein CmAP/MBL-T using the proposed strain.

EFFECT: invention allows obtaining the specified highly purified preparation of a hybrid protein with properties of recombinant high-activity alkaline phosphatase CmAP and highly specific mannan-binding lectin of C-type MBL-T with regard to carbohydrate components of glycoproteins of cancer cells of uterine cervix and can be used for a high-sensitivity lectin-immunoferment method of early differential diagnostics of cervical cancer.

3 cl, 2 dwg, 3 ex

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RU 2 538 169 C1

Authors

Balabanova Larisa Anatol'Evna

Koval'Chuk Svetlana Nikolaevna

Golotin Vasilij Aleksandrovich

Bulgakov Aleksandr Aleksandrovich

Rasskazov Valerij Aleksandrovich

Dates

2015-01-10—Published

2013-09-11—Filed